Ultra-performance Liquid Research Articles

Metabolomics Reveals the Mechanism by Which Sodium Butyrate Promotes the Liver Pentose Phosphate Pathway and Fatty Acid Synthesis in Lactating Goats

This study aimed to explore the effects of sodium butyrate on liver metabolism in goats subjected to a high-concentrate diet. We randomly assigned twelve Saanen-lactating goats into two groups, one of which received a high-concentrate diet (concentrate: forage = 60:40, control group), while the other received the same basal diet supplemented with sodium butyrate (SB) (10 g/kg basal diet, SB group). Compared with the control diet, the SB diet considerably increased the milk fat percentage and content (p < 0.05), with an increase of 0.67% in the milk fat content of the SB group. By employing a global metabolomics approach based on ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS), we identified 6748 ions in ESI+ mode and 3573 ions in ESI− mode after liver isolation from both groups. A total of twenty-three metabolites, including phospholipids, fatty acids, and ribose phosphate, were found to be dysregulated according to a search against the human metabolome database (HMDB). Pathway analysis revealed activation of the pentose phosphate pathway, glycerophospholipid metabolism, and unsaturated fatty acid synthesis. The SB diet also modulated the expression of key lipogenic enzymes, such as acetyl-CoA carboxylase (ACC) and stearoyl-CoA desaturase (SCD-1), which are downstream targets of the transcription factor sterol regulatory element-binding proteins-1c (SREBP-1c), inducing a significant upregulation (p < 0.05). Furthermore, 6-phosphogluconate dehydrogenase (6PGDH) levels in the liver were elevated after the lactating goats were fed the SB diet (p < 0.05). Our study reveals that the SB diet may offer substantial benefits in enhancing the milk quality of subacute ruminal acidosis (SARA) goats. This is accomplished by augmenting the activity of the liver pentose phosphate pathway and the process of de novo fatty acid synthesis in lactating goats.

Exposure to Volatile Organic Compounds and Blood Pressure in NHANES 2011 to 2018

BACKGROUND: Volatile organic compounds (VOCs) are ubiquitous environmental pollutants. Exposure to VOCs is associated with cardiovascular disease risk factors, including elevated blood pressure in susceptible populations. However, research in the general population, particularly among nonsmoking adults, is limited. We hypothesized that higher VOC exposure is associated with higher blood pressure and hypertension, among nonsmokers. METHODS: We included 4 cycles of data (2011–2018) of nonsmoking adults (n=4430) from the National Health and Nutrition Examination Survey. Urinary VOC metabolites were measured by ultraperformance liquid chromatography-mass spectrometry, adjusted for urine dilution, and log-transformed. We estimated mean differences in blood pressure using linear models and prevalence ratio of stage 2 hypertension using modified Poisson models with robust standard errors. Models were adjusted for age, sex, race and ethnicity, education, body mass index, estimated glomerular filtration rate, and National Health and Nutrition Examination Survey cycle. RESULTS: Participants were 54% female, with a median age of 48 years, 32.3% had hypertension, and 7.9% had diabetes. The mean differences (95% CI) in systolic blood pressure were 1.61 (0.07–3.15) and 2.46 (1.01–3.92) mm Hg when comparing the highest with the lowest quartile of urinary acrolein (N-acetyl-S-[2-carboxyethyl]-L-cysteine) and 1,3-butadiene (N-acetyl-S-[3,4-dihydroxybutyl]-L-cysteine) metabolites. The prevalence ratios for hypertension were 1.06 (95% CI, 1.02–1.09) and 1.05 (95% CI, 1.01–1.09) when comparing the highest with lowest quartiles of urinary acrolein (N-acetyl-S-[2-carboxyethyl]-L-cysteine) and 1,3-butadiene (N-acetyl-S-[3,4-dihydroxybutyl]-L-cysteine), respectively. CONCLUSIONS: Exposure to VOCs may be a relevant yet understudied environmental contributor to cardiovascular disease risk in the nonsmoking, US population.

Detection of Ochratoxin A in Maize and Its Potential Impact on Avian Pathology in Romanian Farms

Ochratoxin A (OTA) is a nephrotoxic mycotoxin that commonly contaminates maize, posing significant health risks to both poultry and humans. In this study, a rapid and sensitive method utilizing ultra-performance liquid chromatography coupled with fluorescence detection (UPLC-FLD) was developed for the quantification of OTA levels in maize. The method utilizes immunoaffinity column purification for improved specificity. Accuracy and precision were validated in line with European Union Reference Laboratory (EURL-MP) guidelines, meeting regulatory standards for linearity, trueness, detection and quantification limits, precision, and uncertainty, as per European Commission Regulation (EC) No. 401/2006 and its amendments. The method demonstrated an average recovery rate of 116.78% for maize, with RSDwR values (within-laboratory reproducibility) of 12.72%. Furthermore, OTA occurrence and its possible effects were investigated in several farms in South Romania, where necropsy and histopathological analyses of poultry revealed severe kidney damage, including renal tubular degeneration.

Effect of dietary supplementation of yeast culture Saccharomyces cerevisiae in lactating female goats

This study was designed to investigate the effects of adding a novel yeast culture, Saccharomyces cerevisiae refermented sorghum distiller's dried grains with solubles (SSDDGS), to the diets of lactating female goats on lactation performance and lamb growth performance. We divided 10 lactating Dazu black goats of similar age, weight, and offspring into two groups: one fed a pelleted diet with 50 g/day SSDDGS (ET), and the other without SSDDGS as a control (EC) for 7 weeks. We monitor the weight changes of each goat and collect blood and milk samples from experimental ewes at specific times for hormone and milk composition determination. We use ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) to detect metabolites in the serum of lactating ewes. Our results showed that SSDDGS supplementation significantly reduced female goats' average daily weight loss during weeks 2–4 of lactation and increased serum IGF-1 and prolactin levels at week 4 (p < 0.05). SSDDGS supplementation in early lactation significantly increased milk protein, lactose, and ash content (p < 0.05). UPLC-MS/MS analysis showed that SSDDGS changed the levels of 58 metabolites in the serum of lactating goats. These metabolites were mainly involved in the sohingolipid signaling pathway, and cysteine, methionine, and sphingolipid metabolism. In summary, Yeast culture SSDDGS reduced weight loss, enhanced milk quality, and modified metabolic profiles in early lactation goats, providing insight into the potential regulatory role and mechanism of yeast culture in lactation female goats.

Integrative Transcriptomic and Metabolomic Analysis Reveals Quinoa Leaf Response Mechanisms to Different Phosphorus Concentrations During Filling Stage

Quinoa is an annual self-pollinating plant rich in polyphenols, flavonoids, saponins, and amino acids; its protein balance closely aligns with the ideal recommendation set by the Food and Agriculture Organization. Therefore, quinoa is considered the most suitable “all-nutrient food”. Phosphorus fertilization plays an important role in restricting the growth and development of quinoa; however, the effects of phosphorus fertilizer on quinoa growth remain unclear. Therefore, we conducted metabolome and transcriptome analyses on quinoa leaves during the filling stage, subjecting plants to different doses of phosphorus fertilizer. Overall, phosphorus treatment exerted a significant impact on the phenotypic characteristics of quinoa. Specifically, through a combined analysis of ultra-performance liquid chromatography–tandem mass spectrometry and transcriptome analysis, we identified the alteration and regulation of specific metabolites and genes within flavonoid biosynthesis pathways; this comprehensive evaluation helped elucidate the response mechanism of quinoa leaves during the grouting stage under various phosphorus conditions. Ultimately, the results of this study provide a reference for the selection of quinoa cultivars that exhibit tolerance to low- or high-phosphorus stress; additionally, we offer a theoretical basis for the rational application of phosphorus fertilizer and the enhancement of phosphorus utilization efficiency.

Impact of Drying on Phytonutritional Compounds, In Vitro Antioxidant Activity and Cytotoxicity of Spiny Saltbush (Rhagodia spinescens)

The Spiny saltbush (Rhagodia spinscens) is a halophyte species with the potential to provide natural ingredients used in food and pharmaceutical industries. In food and pharmaceutical applications, drying is necessary to maintain shelf-life, which reduces phytonutrient content. In this study, changes in the nutritional composition, phenolic and carotenoid profiles of radical antioxidant scavenging activity [(2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS)], antioxidant power [ferric reducing antioxidant ability assay (FRAP)], and cytotoxicity of freeze- and oven-dried (55 °C for 24 h) spiny saltbush were determined. Sodium (4.72 g/100 g dry weight (DW), potassium (6.86 g/100 g DW), calcium (4.06 g/100 g DW), zinc (372 mg/kg DW) and protein content were higher in oven-dried samples than freeze-dried samples. Ultra-performance liquid chromatography–mass spectrometry analysis detected 18 metabolites in saltbush extracts. Partial Least-Squares Discriminant Analysis, Hierarchical Cluster Analysis, and Variable Importance in Projection discriminated between freeze-dried and oven-dried samples. Freeze-dried samples retained more individual metabolites than oven-dried samples, while oven-dried samples had higher antioxidant activity (ABTS and FRAP), lutein, trans-β carotene, and cis-β-carotene. Correlation analysis identified potential antioxidant candidates between phenolic and carotenoid compounds. Neither freeze-dried nor oven-dried spiny saltbush samples showed cytotoxicity. The study uncovered changes in phytonutritional compounds after the oven and freeze-drying spiny saltbush, a potential salt alternative and functional ingredient for the food industry.

Antibiotic exposure and risk of overweight/obesity in children: a biomonitoring-based study from eastern Jiangsu, China

ObjectiveTo describe antibiotic exposure in children and explore its association with overweight/obesity.MethodsIn June 2022, 328 kindergarten and primary school children were selected from Nantong city in Jiangsu Province. Questionnaires were distributed, and morning urine samples were obtained. Total urinary concentrations of 41 antibiotics were measured using ultra-performance liquid chromatography and tandem mass spectrometry. The rates of antibiotic exposure were expressed as percentages (%), specific percentiles (P95 and P99), and the maximum values were used to describe the concentration of antibiotics. The association between urinary antibiotic creatinine-adjusted and overweight/obesity was analyzed using logistic regression.ResultsA total of 328 children were initially recruited, of which 295 aged 3–8 years met the inclusion criteria and were finally included in the study. The biomonitoring results revealed that 35 antibiotics were detected, with a total detection frequency of 98.31%. Among the included children, 24.75% were classified as overweight/obesity. Multinomial logistic regression analyses revealed significant associations between overweight/obese and exposure to veterinary antibiotics (VAs) and preferred veterinary antibiotics (PVAs). After adjusting for various overweight/obesity-relevant variables, higher exposure to sulfamethoxazole [OR = 2.35, 95% confidence interval (CI):1.17–4.70], norfloxacin (OR = 2.66, 95% CI: 1.01–7.08), and fluoroquinolones (OR = 1.97, 95% CI: 1.02–3.78) were significantly associated with overweight/obesity (p < 0.05). In addition, after stratification by sex and adjustment for confounding variables, sex-specific differences were observed in the association between antibiotic exposure and overweight/obesity. Notably, these associations were predominantly observed among boys.ConclusionChildren were extensively exposed to antibiotics. Exposure to certain types of veterinary antibiotics and preferred veterinary antibiotic exposure, mainly through food or drinking water, are associated with an increased risk of overweight/obesity in children.

Implemention of Innovative Process Analytical Technologies to Characterize Critical Quality Attributes of Co-Formulated Monoclonal Antibody Products.

Characterizing co-formulated monoclonal antibodies (mAbs) poses significant challenges in the pharmaceutical industry. Due to the high structural similarity of the mAbs, traditional analytical methods, compounded by the lengthy method development process, hinder product development and manufacturing efficiency. There is increasing critical need in the pharmaceutical industry to streamline analytical approaches, minimizing time and resources, ensuring a rapid clinical entry and cost-effective manufacturing. This study investigates the application of process analytical technologies (PAT) to address such challenges. Our investigation introduces two complementary technologies, on-line ultra-performance liquid chromatography (online UPLC) and multimode fluorescence spectroscopy (MMFS), as potential PAT tools tailored for characterizing critical quality attributes (CQA) in co-formulated mAb products. Specifically, the CQAs under evaluation include the total protein concentration of the mAbs within the co-formulation and the ratio of mAb A to mAb B. Online UPLC enables direct and automated measurement of the CQAs through physical separation, while MMFS determines them in a non-destructive and more swift manner based on chemometric modeling. We demonstrate these technologies' comparable performance to conventional methods, alongside substantial benefits such as reduced analytical turnaround time and decreased laboratory efforts. Ultimately, integrating them as innovative PAT tools expedites the delivery of therapeutic solutions to patients and enhances manufacturing efficiency, aligning with the imperative for swift translation of scientific discoveries into clinical benefits.

Preparation, characterization and mechanism of SiO2@SiO2 core-shell microspheres through polymerization-induced colloid aggregation for fast separation using ultra performance liquid chromatography

The polymerization-induced colloid aggregation (PICA) method is commonly used to create SiO2@SiO2 core–shell silica microspheres (CSSMs), but it often encounters challenges such as incomplete shell coating and poor reproducibility. In this paper, the formation mechanism of the silica shell layer during the preparation of SiO2@SiO2 CSSMs using the PICA method was investigated. It was found that ureido modification can reduce the Zeta potential of the silica core surface, facilitating the deposition of coacervates formed by urea-formaldehyde resin (UF) and silica nanoparticles on the silica core surface to form the SiO2 shell layer when the Zeta potential of the surface is in the range of −20.1 mV to −4.8 mV. By controlling the interfacial state and surface potential of the SiO2 core, the issue of inconsistent reproducibility in preparing SiO2@SiO2 CSSMs can be overcome. Furthermore, optimization of experimental parameters such as pH, reaction temperature, water content, and reaction time can enhance the formation process. The thickness of the shell and pore size of CSSMs can be successfully adjusted by varying the reaction time and the particle size of colloidal silica sol, respectively. With optimal conditions, the semi-scale production yield of SiO2@SiO2 CSSMs can be increased to 50 g. The SiO2@SiO2 CSSMs were modified with octadecyltrichlorosilane (ODS) to be used as stationary phases in reversed phase liquid chromatography (RPLC) for fast separation of small solutes, peptides, and proteins. The superior separation efficiency indicates that the improved PICA method has the potential to be utilized as an alternative to the layer-by-layer method for large-scale production of SiO2@SiO2 CSSMs.

Phenolic Compound Profiles, Antioxidant, and Cytoprotective Activity of Elaeagnus conferta Roxb. Fruit.

In this paper, three varieties of Elaeagnus conferta Roxb fruits prepared by ultrasonic-assisted extraction from a subtropical region southwest of China were utilized as raw materials to investigate their phenolic profiles, antioxidant activities, and protective effects on injured human umbilical vein endothelial cells (HUVECs). The ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) findings revealed that fifteen substances, including seven phenolic acids, seven flavonoids, and one gallic acid derivative, were discovered. The dihydromyricetin, ellagic acid, gallic acid were the predominant phenolic compounds in all E.conferta fruits. These E.conferta fruits extracts shown excellent antioxidant activity varied from 2.258±0.03~7.844±0.39 μM Trolox/g and protective effect on HUVECs injured by H2O2 through decrease the level of ROS, MDA, LDH and enhance the SOD level. These finding indicate that E.conferta is a valuable source of high-capacity antioxidants that might be used as an alternative material for food industries.

Genetic and QTL analysis of sugars and acids content in sweet cherry (Prunus avium L.)

Abstract Sweet cherry is very appreciated by consumers because of its attractive appearance and taste, which is determined by the balanced sweet-sour flavor. In this work, the genetics of soluble solid content (SSC), titratable acidity (TA), sugars and organic acids was investigated in sweet cherry to facilitate breeding improvement for fruit quality. The fruits of five sweet cherry populations (N = 372), three F1 and two F2, were sampled over two years to evaluate SSC, TA, and the content of individual sugars (glucose, fructose, sorbitol, and sucrose) and organic acids (malic, quinic, oxalic, citric and shikimic) by ultra-performance liquid chromatography (UPLC). Glucose, followed by fructose, was the most abundant sugar, while malic acid was the predominant acid. Sorbitol and malic acid were the most stable compounds between years, and had the highest heritability, being also the best correlated to SSC and TA respectively, revealing their relevance for breeding. Significantly positive correlations were observed among sugars and SSC, and acids and TA, but high interannual variability between years was observed for all traits. QTL mapping for SSC, sugars, TA, and organic acids was performed using a multi-family approach with FlexQTL™. Twenty QTLs were detected consistently during the two phenotyped years, and several relevant regions with overlapping QTLs for sugars and acids were also identified. The results confirmed major stable SSC and TA QTLs on the linkage groups 4 and 6, respectively. Within the main LG4 SSC QTL region, where maturity and fruit development time QTLs have been previously detected, three stable sugar (glucose, sorbitol, and sucrose) and two acid (quinic, shikimic) QTLs were also identified, suggesting a pleiotropic effect of ripening date on the content of these compounds. The major malic acid QTL overlapped with TA QTL on LG6, thus TA QTL mapping in LG6 may correspond to malic acid QTLs. Haplotype analyses of major SSC and sugars QTL in LG4, and TA and malic acid in LG6 revealed haplotypes of breeding interest. Several candidate genes previously identified in other Prunus fruit species, like peach, were found to collocate with the QTLs detected herein. This work reports QTLs regions and haplotypes of sugar and acid content in a Prunus non-climacteric stone fruit for the first time.

Antibiotic contaminants and their impact in Gingee River, Puducherry: insights from SPE-UPLC–MS/MS and zebrafish study

Abstract The accumulation of antibiotic residues in ecosystems is intricately tied to the proliferation of bacterial resistance to antibiotics, with far-reaching consequences for the health and welfare of both humans and animal well-being. The analytical approach integrates solid phase extraction (SPE) with ultra performance liquid chromatography-mass spectrometry (UPLC–MS/MS) for quantification of multiclass antibiotic residues. Upon applying the aforementioned method to analyse water samples collected from the Gingee River, revealed the existence of five distinct antibiotics. This is the first study reporting antibiotic concentration in the Gingee River, Puducherry. The concentrations of nalidixic acid, sulfamethoxazole, and tetracycline were determined to be 8.5, 6.9, and 4.8 μg/L, respectively. Metronidazole and trimethoprim were detected at concentrations below the quantifiable limit. The microbial study of water samples also indicated that Shigella sp. and Acinetobacter sp. were the most predominant bacterial species present. Our preliminary observation underscores the importance of comprehending the intricate relationship between the presence of antibiotics in water and the concurrent proliferation of antibiotic-resistant bacteria within bacterial populations in the Gingee River. Further, we evaluated the developmental toxicity of environmentally relevant concentrations of antibiotics in zebrafish. The zebrafish model confirms that these antibiotics are sublethally hazardous to human health at environmentally relevant concentrations. This integrated approach allows unique views on the environmental impact of antibiotic residues, their role in the evolution of antibiotic resistance, and their impact on human health.

Development and validation of an LC‒MS/MS method for the determination of cyclocreatine phosphate and its related endogenous biomolecules in rat heart tissues

The cardioprotective drug cyclocreatine phosphate has been awarded Food and Drug Administration-orphan drug designation for the prevention of ischemic injury to enhance cardiac graft recovery and survival in heart transplantation. Cyclocreatine phosphate is the water-soluble derivative of cyclocreatine. Estimating the levels of Cyclocreatine phosphate, Adenosine triphosphate, Creatine Phosphate, Creatine and Cyclocreatine helps us in understanding the energy state as well as evaluating the heart cells’ function. The quantification of endogenous compounds imposes a challenging task for analysts because of the absence of a true blank matrix, whose use is required according to international guidelines. Recently, the International Council for Harmonization issued a new guideline that contains guidance on the validation of methods used to quantify endogenous components, such as the background subtraction approach that was employed in our current study. Specifically, we developed and validated a sensitive, reliable and accurate liquid chromatography-tandem mass spectrometry assay to determine simultaneously the levels of mentioned endogenous compounds in rat heart tissue. Tissue samples were prepared by protein precipitation extraction using water: methanol (1:1). Using Ultra Performance Liquid Chromatography, Chromatographic separation was achieved with ZORBAX Eclipse Plus C18 4.6 × 100 mm,3.5 μm column and conditions as following: ammonium acetate (pH 8.5): acetonitrile, 70:30 mobile phase, 0.7 mL/min flow rate and 25 °C temperature. Electrospray ionization mass detector with Multiple reaction monitoring mode was then employed, using both positive and negative modes, Analysis was carried out using 5.00–2000.00 ng/mL linear concentration range within 2 min for each analyte. According to Food and Drug Administration guidelines for bioanalytical methods, validation was carried out. We investigated the matrix effect, recovery efficiency and process efficiency for the analyte in neat solvent, postextraction matrix and tissue. The results stated mean percentage recoveries higher than 99%, accuracy 93.32–111.99%, and Relative Standard Deviation (RSD) below 15% within the concentration range of our study which indicated that target analytes’ stability in their real matrix is sufficient under the employed experimental conditions.

Evaluation of protein impurities in Ademetionine 1,4-Butanedisulfonate

Ademetionine 1,4-Butanedisulfonate (SAMe) is widely used as a prescription drug to treat cholestasis associated with liver disease, and is also used to treat depression, Alzheimer's disease and other diseases. Currently, the main way to produce SAMe is by fermentation of S-adenosylmethionine synthetase（SAMS）. However, during the fermentation process, host cells produce contaminants unrelated to the treatment. Among them, host cell protein (HCP), which is co-purified with the drug, is the main impurity in the production process. HCP-induced antigenic reactions can lead to allergies or other adverse reactions and affect drug quality, efficacy, and safety. Enzyme-linked immunosorbent assay (ELISA) and mass spectrometry (MS) are currently used as the main analytical methods for measuring HCP. The ELISA method may ignore some HCPs with low immunogenicity, while mass spectrometry is primarily employed for the characterization of proteomes. Orthogonal methods can more effectively evaluate impurities in drugs. In this study, ELISA was initially utilized to quantify the content of Saccharomyces cerevisiae host protein in 11 batches of SAMe active pharmaceutical ingredient (API) from two suppliers, then the above APIs were qualitatively analyzed by MS method. Following the identification of surrogate peptide of SAMS, an Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry（UPLC-MS/MS）method was established and validated, the SAMS protein residues in 11 batches of samples were subsequently determined. The results demonstrated that the levels of Saccharomyces cerevisiae host protein and SAMS in the samples were both low, with values below 20 ppm and 1.19 ppm, respectively. The qualitative results also indicate that the types of peptide residues in the samples are more diverse than those of protein residues. Furthermore, residues of proteins and peptides can be detected in all samples, which underscores the importance of evaluating the HCP in API. This study employs a range of complementary detection methods to conduct a comprehensive and sensitive evaluation of total HCP and specific proteins in drugs, thereby guiding more informed assessments of the risks posed by HCP impurities in raw materials during pharmaceutical processes.

An Integrated Approach to Identify the Q-Markers of Banxia-Houpo Decoction Based on Nontargeted Multicomponent Profiling, Network Pharmacology, and Chemometrics.

The inherent complexity of traditional Chinese medicine (TCM) poses significant challenges in directly correlating quality evaluation with clinical efficacy. Banxia-Houpo Decoction (BHD), a classical TCM formula, has demonstrated efficacy in treating globus hystericus. However, the intricate composition of BHD, which contains both volatile and non-volatile active components, complicates efforts to ensure its consistent quality and clinical effectiveness. The aim of this study was to introduce an integrated approach that combines non-targeted multicomponent analysis, network pharmacology, and multivariate chemometrics to identify quality markers for the effective quality control of BHD. First, a nontargeted high-definition MSE method based on ultraperformance liquid chromatography-quadrupole time-of-flight-mass spectrometry (UHPLC-QTOF-MS) was developed for the comprehensive multi-component characterization of BHD. Next, the quality markers of nonvolatile compounds in BHD were identified through network pharmacology analysis. Subsequently, volatile organic compounds (VOCs) in BHD samples were analyzed via headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) and headspace gas chromatography-ion mobility spectrometry (HS-GC-IMS). Finally, the orthogonal partial least squares discriminant analysis (OPLS-DA) model was applied to screen for potential markers. Based on in-house library-driven automated peak annotation and comparison with 25 reference compounds, 128 components were identified for the first time. Additionally, honokiol, magnolol, magnoflorine, 6-gingerol, rosmarinic acid, and adenosine were preliminarily identified as potential quality markers for BHD through network pharmacology analysis. By employing two complementary techniques, HS-SPME-GC-MS and HS-GC-IMS, a total of 145 volatile compounds was identified in the BHD samples. Four potential differential VOCs in the BHD samples were further identified based on the variable importance in projection (VIP ≥ 1.5) using HS-GC-IMS combined with chemometric analysis. In conclusion, this study not only contributes to establishing quality standards for BHD but also offers new insights into quality assessment and identification in the development of classical formulations enriched with volatile components.

Differences in IgG afucosylation between groups with and without carotid atherosclerosis

BackgroundA previous study demonstrated that N-glycosylation profiles of IgG are associated with subclinical atherosclerosis in a British population. However, the generalisability of this finding to other ethnic groups remains to be investigated, and it has yet to account for additional traditional atherosclerotic risk factors. The present study, thus, aims to explore IgG N-glycosylation profiles in Han Chinese with atherosclerosis, and their potential role in atherosclerosis, while controlling for traditional atherosclerotic risk factors.MethodsData of this case-control study were obtained from an established umbrella Health Examination Cohort Study (registration number: ChiCTR2100048740). The investigation was conducted at the Health Care Centre of the First Affiliated Hospital of Shantou University Medical College in China, from August 1, 2021, to July 31, 2022. A sample of 69 carotid atherosclerosis (CAS) cases was recruited from the umbrella cohort, along with 69 controls without carotid atherosclerosis, matched by traditional atherosclerosis-related risk factors, including gender, age, smoking, alcohol consumption, hypertension, diabetes, dyslipidemia and obesity. Subsequently, serum IgG N-glycosylation was profiled using Ultra-Performance Liquid Chromatography.ResultsAfter propensity score matching, the relative abundance of IgG fucosylation in CAS cases was significantly lower than that in controls [95.32 (92.96, 95.99) vs. 95.96 (94.70, 96.58), P = 0.022]. The traditional atherosclerosis-related risk factors showed no statistically significant difference between CAS cases and controls (P > 0.05).ConclusionsThe reduced fucosylation of IgG in CAS cases underscores the pivotal role of afucosylation in CAS. Enhancing the inflammatory capability of IgG via initiating antibody-dependent cell-mediated cytotoxicity could be the potential mechanism behind this, which should be further verified by functional studies.

Using metabolomics to explore the effects of epigenetic-modification strategies on the metabolites of Acanthus ilicifolius L. endophytic fungi against ovarian cancer

Ovarian cancer is a serious threat to women's health and safety. So far, people have discovered more than 130 small molecule compounds of natural origin for anti-tumor, of which approximately 50% are of microbial origin. The Acanthus ilicifolius L. species is primarily distributed in the Guangdong, Hainan, and Guangxi regions of China and grows in tidally accessible coastal areas. Recent studies have revealed that Acanthus ilicifolius L. extracts are endowed with a range of pharmacological properties, including anti-inflammatory, hepatoprotective, antioxidant, and antitumor activities. Endophytic fungi are commonly found in the healthy tissue and organs of medicinal plants. These fungi and the plants they inhabit form mutually beneficial symbiotic relationships. Endophytic fungi produce a series of secondary metabolites, with active substances having shown great economic value and applications prospects in drug research and development as well as for the biological control of plant diseases. Secondary metabolites production by endophytic fungi is regulated by specific gene clusters, and several techniques have been used to stimulate the secondary metabolic processes of fungi, including epigenetic-modification and OSMAC (one strain many compounds) strategies, co-culturing, and gene modification. Among these, epigenetic modification has been shown to be effective; this strategy involves the addition of small-molecule epigenetic modifiers to the culture medium, thereby activating silenced biosynthetic gene clusters without altering the DNA sequences of the fungi. This approach facilitates the expression of silenced genes in endophytic fungi, thereby increasing the number and diversity of secondary metabolites. Furthermore, it assists in overcoming the inhibition of microbial secondary-metabolite synthesis under laboratory conditions, and enhances silenced-gene expressions. The advent of novel analytical techniques and bioinformatics has provided a comprehensive, multifaceted, and holistic understanding of fungal metabolism through the development of metabolomics as a research platform. However, few studies have combined anti-ovarian cancer-activity screening with metabolomic approaches in the search for activity-differentiating metabolites from endophytic fungi under the intervention of epigenetic modifiers. Herein, we investigated the impact of epigenetic modifiers on the secondary metabolites of the endophytic Diaporthe goulteri fungus from Acanthus ilicifolius L. to determine their potential anti-ovarian cancer activities. Crude extracts were obtained by controlling three variables: the number of fermentation days, the type of epigenetic modifier, and its concentration, with activities screened using the CCK-8 (cell counting kit-8) method. Ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was subsequently employed for non-targeted metabolomic analysis. A multivariate statistical analysis model was constructed using principal component analysis and orthogonal partial least squares-discriminant analysis, which combines model and variable importance projection, with qualitative screening performed and significant changes (variable importance in the projection (VIP)≥1; P<0. 05) determined. Fifteen differential metabolites were identified in the fungal and epigenetic modification group, primarily comprising polyketides, amino acids, derivatives, alkaloids, and organic acids, including prenderol, glycine, valine, 2-ethylcaproic acid, rubratoxin B, finasteride, 6-silaspiro[5.5]undecane, 1-(2-nitrophenoxy)octane, heptadecene, 1-pentadecene, 11-ketoetiocholanolone, 3-(1-ethyl-1,3,3-trimethyl-2,3-dihydro-1H-inden-5-yl)butanal, N2-benzoylarginine, tabutrex, (3aR,6S,6aS)-6-(4-hydroxy-2-methoxy-2-butanyl)-4,4-dimethylhexahydro-1(2H)-pentalenone, and 8-aminoquinoline. The expressions of prenderol, 1-(2-nitrophenoxy)octane, 3-(1-ethyl-1,3,3-trimethyl-2,3-dihydro-1H-inden-5-yl)butanal, N2-benzoylarginine, and 8-aminoquinoline were downregulated, whereas the expressions of the remaining 10 substances were upregulated. Polyketides were the main components that exhibited higher expressions. This study showed that latent active differential metabolites can be searched by combining anti-ovarian cancer-activity screening with metabolomics analysis, thereby providing a reference for the further development of Acanthus ilicifolius L. resources and the subsequent targeted isolation of active compounds.

Screening and Identification Quality Markers for Antioxidant Activity ofTetrastigma hemsleyanumfrom Different Origins using Spectrum-effect Correlation Analysis

Background::Tetrastigma hemsleyanum (THL) is a traditional medicinal plant with various biological activities, it has been validated that several THL extracts have strong pharmacological activities. Until now, there is no complete and systematic germplasm identification and quality evaluation standard.Objective::Establishing fingerprint spectra of THL from different sources, evaluating their antioxidant activity and identifying quality markers.Methods::In this paper, establishing fingerprints and identifying quality markers through UPLC-QTOF- MS/MS. The antioxidant capacity of forty-one batches of THL from multiple regions was determined by 1,1-Diphenyl-2-picrylhydrazyl radical (DPPH), 2,2'-Azinobis-(3-ethylbenzthiazoline-6- sulphonate) (ABTS), and Ferric reducing antioxidant power (FRAP) assays.Results::The results indicated that THL methanol extracts showed good antioxidative activity and eight THL samples, including THL aerial parts, showed high antioxidative activity. Besides, Ultra Performance Liquid Chromatography (UPLC) fingerprints of THL from different places were established and chromatographic peaks probably correlated with antioxidant capacity were screened by principal component analysis (PCA) and partial least square regression (PLS) analysis.Conclusion::The chemical constituents of these peaks were identified as possible quality markers for THL quality evaluation. In this study, twelve effective substances were preliminarily determined, such as isovitexin, L-epicatechin, procyanidin B, sitosterol, etc.

Quantitative Lipidomics Reveals the Effects of Roasting Degree on Arabica Coffee Beans Lipid Profiles

Lipids in coffee beans undergo morphological changes and migration during the roasting process, but the impact of roasting intensity on lipid profiles remains unclear. The lipidomics characteristics of coffee beans with different degrees of roasting (unroasted green beans, light roast, medium roast, and dark roast) were compared using ultra performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS). A total of 571 lipid metabolites were identified, encompassing 25 species of fatty acyls (FA), 142 species of glycerophospholipids (GP) across 12 subclasses, 42 species of sphingolipids (SP) across four subclasses, 360 species of glycerolipids (GL) across ten subclasses, and two species of prenol lipids (PR). GL content was found to be the highest, followed by FA and GP. Through multivariate analysis, 160 lipids were identified that exhibited significant differences among beans with different degrees of roasting. GP content was significantly influenced by the degree of roasting (p<0.05), with diverse dynamic evolution observed through K-means cluster analysis. These findings have important implications for understanding lipid transformation and its effect on the flavor profile during coffee roasting.

Simultaneous determination of 17 perfluoroalkyl substances in beef by EMR-Lipid dispersive solid-phase extraction and ultra performance liquid chromatography-tandem mass spectrometry

The ability to accurately analyze perfluoroalkyl substance (PFAS) levels in beef is imperative in order to effectively assess food-safety risks and ensure consumer safety because PFASs are harmful and prevalent in beef. In this study, we developed a rapid and accurate method for the simultaneously determination of the 17 PFASs in beef using dispersive solid-phase extraction (d-SPE) and ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), and optimized the mobile phase system, extraction solvent, and d-SPE materials. Samples were finally extracted using 0.1% (v/v) formic acid in acetonitrile, cleaned using d-SPE with PSA, C18, GCB, and EMR-Lipid, separated using an Acquity Premier BEH C18 column (100 mm×2.1 mm, 1.7 μm) with 0.5 mmol/L ammonium fluoride aqueous solution and methanol as the mobile phases at a flow rate of 0.3 mL/min. Analytes were detected in negative ion switching mode (ESI-) with multiple reaction monitoring (MRM) scanning, and quantitatively analyzed using the internal standard method. The 17 PFASs exhibited linearity in the 0.2-20.0 μg/L range under the optimal experimental conditions, with correlation coefficients of 0.9915-0.9999. The method delivered limits of detection (LODs) of 0.003-0.007 μg/kg and limits of quantification (LOQs) of 0.01-0.02 μg/kg. The 17 PFASs exhibited recoveries of 71.1%-127.4% with RSDs of 0.6%-14.4% when spiked at three levels (0.05, 0.5, and 1.8 μg/kg). We optimized the mobile phase system, which revealed that, compared with 2.0, 5.0, and 10.0 mmol/L ammonium formate or ammonium acetate in aqueous methanol, 0.5 mmol/L ammonium fluoride in aqueous methanol exhibited higher sensitivities for all the 17 PFASs, with PFASs bearing long-chain carboxylic acids (C10-C18) showing 1-2 fold increases in sensitivity. PFASs do not dissociate in acidic environments, favoring their entry into the organic phase. Therefore, we investigated the effect of extractant acidity, which revealed that the 17 PFASs were better extracted using 0.1% (v/v) formic acid in acetonitrile. The beef matrix has a complex composition; consequently, d-SPE adsorbents were required to purify samples and reduce matrix effects. The purification effects of four adsorbents (PSA, C18, GCB, and EMR-Lipid) toward the 17 PFASs and the amount of EMR-Lipid used were investigated, which revealed that 100 mg PSA+80 mg C18+40 mg GCB+150 mg EMR-Lipid exhibited superior matrix-purification behavior. We also investigated the effects of various injection solutions and types of syringe filter, with pure methanol selected for reconstitution and high-speed supernatant centrifugation applied prior to injection. The developed method is simple, rapid, sensitive, and reproducible, and can be used to simultaneously, rapidly, and accurately determine various perfluoroalkyl compounds in beef.