Ultra-high Performance Liquid Chromatography-quadrupole Time-of-flight Mass Spectrometry Research Articles

Ultra-High-Performance Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry for Simultaneous Pesticide Analysis and Method Validation in Sweet Pepper.

Pesticides effectively reduce the population of various pests that harm crops and increase productivity, but leave residues that adversely affect health and the environment. Here, a simultaneous multicomponent analysis method based on ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) pretreated by the QuEChERS method was developed to control the maximum residual levels. Among the 140 pesticides with high frequency of detection in agricultural products in Gyeongnam region in Korea for 5 years, 12 pesticides with high detection frequency in sweet pepper were selected. The analytical method is validated, linearities are r2 > 0.999, limit of detection (LOD) ranges from 1.4 to 3.2 µg/kg, and limit of quantification (LOQ) ranges from 4.1 to 9.7 µg/kg, and the recovery rate was 81.7-99.7%. In addition, it was confirmed that a meaningful value of these parameters can be achieved by determining the measurement uncertainty. The results proved that parameters such as recovery rate and relative standard deviation of the analysis method were within international standards. Using the developed method, better and safer sweet peppers will be provided to consumers, and effective pesticide residue management will be possible by expanding to other agricultural products.

Rapid screening of active ingredients and action mechanisms of Ecliptae Herba for treating Alzheimer's disease by UPLC-Q-TOF/MS and “component-target-pathway” network

Alzheimer's disease (AD) is a common neurodegenerative disease. The drugs widely used in clinic are mainly single-target drugs for symptomatic treatment, which can only alleviate symptoms to a certain extent. Ecliptae Herba (EH) is considered a potential therapeutic drug for AD due to its neuroprotective effects. Although EH has a clear anti-AD effect, the material basis and mechanism remain unclear. Therefore, we adopted an efficient analytical technique, namely ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS), combined with “component-target-pathway” network to explore the active components and potential mechanisms of EH in treating AD. Due to the high sensitivity of UPLC-Q-TOF/MS, a total of 50 components were identified in EH. Among them, 20 and 12 compounds were found in plasma and brain samples, respectively. The network pharmacology analysis revealed that apigenin, luteolin, ecliptasaponin A, chlorogenic acid, wedelolactone, and quercetin were the active components, which could affect the serotonergic synapse, calcium and cAMP signaling pathways by regulating related targets such as EGFR, PRKCA, BRAF and ERBB2. This study clarified that EH can exert anti-AD effect through multi-component, multi-target and multi-pathway characteristics. Furthermore, it offers a good foundation for further in-depth research on the anti-AD effects of EH, and provides a valuable approach for the rapid screening of active components and potential mechanisms of other medicinal plants, potentially bringing changes to the discovery and development of novel therapeutics for neurodegenerative disorders.

Determination of multiclass contaminants in chilli powder based on magnetic multiwalled carbon nanotubes and UPLC-QTOF/MS

A multiclass analysis approach was developed using magnetic multiwalled carbon nanotubes sorbents and ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) for the rapid screening and simultaneous determination of 216 contaminants including 15 mycotoxins, 9 synthetic colourants, and 192 pesticides in chilli powder. The sample preparation process was optimized. The optimal preparation procedure utilized NaCl and NaAc as the salting-out agents, and Fe3O4-MWCNTs as the sorbents, resulting in reduced chemical consumption, improved cleaning performance, and facilitated high-throughput analysis. The proposed method was validated, and satisfactory parameters were obtained. Approximately 85.6% of the target analytes exhibited a weak matrix effect, with the matrix effects falling within the range of 0.8 ∼ 1.2. The method demonstrated acceptable recoveries of the analytes, falling within the range of 62.14%∼119.76% at three fortified levels with relative standard deviations (RSDs) of less than 20%. Additionally, the method’s limit of quantification (LOQ) ranged from ranged from 0.50 μg·kg−1 to 49.56 μg·kg−1. The method was further applied for analysis of 27 chilli powder samples, demonstrating its potential for screening and quantification of multiclass contaminants for spices.

Comparative untargeted metabolomics analysis of serum metabolic alterations in patients infected with hepatitis B virus genotypes B and C

The clinicopathological characteristics and outcomes of patients infected with hepatitis B virus (HBV) differ between genotypes B and C. However, the potential metabolic mechanisms and differences in certain biological pathways associated with different HBV genotypes remain unclear to date. To obtain a better understanding of the effects of liver diseases caused by HBV genotypes B and C on host metabolism, an untargeted metabolomics analysis was performed to assess the differences among the serum metabolic profiles of healthy controls and patients infected with genotypes B or C. Here, a total of 54 serum samples were obtained from healthy controls and patients with chronic HBV infections caused by genotypes B and C, and the samples were subjected to metabolomics analysis. The serum metabolite profiles were analyzed using an ultrahigh-performance liquid chromatography-quadrupole-time-of-flight/mass spectrometry (UHPLC-Q-TOF/MS)-based untargeted metabolomics approach for identifying the differentially expressed metabolites that were upregulated or downregulated by at least 1.5-foldin the serum samples of patients infected with HBV genotypes B and C. A total of 63 metabolites were found to be differentially expressed between the serum samples of healthy controls and patients infected with HBV genotype B, while 57 metabolites were differentially expressed between the serum samples of healthy controls and patients infected with HBV genotype C. The majority of these metabolites were involved in regulating the bile acid (BA) metabolism pathways. The results also indicated that 8 differentially expressed metabolites can serve as potential metabolic biomarkers for distinguishing between individuals infected with the HBV genotype B and C. Altogether, the untargeted metabolomics analysis revealed that infection with HBV genotypes B and C is associated with different metabolic profiles, and the study provided fundamental information for further detailed investigations of the mechanism underlying metabolic dysregulation in HBV infections.

Qualitative and quantitative study of flavonoids in Notoginseng Radix et Rhizoma based on UPLC-Q-TOF-MS and HPLC-DAD

The flavonoids in Panax notoginseng were qualitatively analyzed by ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF-MS), and the content of three main flavonoids in P. notoginseng of different specifications and grades collected from different habitats was determined by HPLC-DAD. Flavonoids and anthocyanins were analyzed by UPLC-Q-TOF-MS/MS in the positive and negative ion modes, respectively. Twelve flavonoid glycosides and one anthocyanin glycoside in P. notoginseng were identified, but no flavonoid aglycones were detected. Among them, 12 compounds were identified in the underground part of P. notoginseng for the first time and eight compounds were first reported in this plant. Moreover, six and four compounds were identified in the Panax genus and the Araliaceae family for the first time, respectively. A method for simultaneous determination of three flavonoids in P. notoginseng was established by HPLC-DAD. The content of flavonoids in 721 P. notoginseng samples of 124 specifications and grades collected from 20 different habitats was simultaneously determined. Among three flavonoids determined, the content of quercetin-3-O-(2″-β-D-xylosyl)-β-D-galactoside was the highest with the average content in the tested samples of 161.0 μg·g~(-1). The content of compounds quercetin-3-O-hexosyl-hexoside and kaempferol-3-O-pentosyl-hexoside was relatively low, with the average content of 18.5 μg·g~(-1)(calculated as quercetin-3-O-sophoroside) and 49.4 μg·g~(-1)(calculated as kaempferol-3-O-sangbu diglycoside). There were significant differences in flavonoids content of samples from different production area. The content of flavonoids in spring P. notoginseng was significantly lower than that in winter P. notoginseng when the other influencing factors such as production areas, germplasm resources, and cultivation conditions were fixed. As for P. notoginseng of different specifications, the flavonoid content in the part connecting the taproot and the aboveground stem was significantly higher than that in other parts. The results of large-scale data showed that the flavonoid content gradually increased with the increase in the number of heads. There were significant differences between the flavonoid content in most specifications and grades, especially the 20-head P. notoginseng and countless head P. notoginseng, whose content was significantly lower and significantly higher than that of other specifications and grades, respectively. This study provides a scientific basis for the study of the effective components and quality control of P. notoginseng from the perspective of flavonoids.

Characterization and identification of chemical components in traditional Chinese medicine Psoraleae Fructus based on UHPLC-Q-TOF-MS

This study was designed to comprehensively characterize and identify the chemical components in traditional Chinese medicine Psoraleae Fructus by establishing an ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry(UHPLC-Q-TOF-MS) method in combination with in-house library. The chromatographic separation conditions(stationary phase, column temperature, mobile phase, and elution gradient) and key MS monitoring parameters(capillary voltage, nozzle voltage, and fragmentor) were sequentially optimized via single-factor experiments. A BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) was finally adopted, with the mobile phase consisting of 0.1% formic acid in water(A) and acetonitrile(B) at the flow rate of 0.4 mL·min~(-1) and column temperature of 30 ℃. Auto MS/MS was utilized for data acquisition in both positive and negative ion modes. By comparison with reference compounds, analysis of the MS~2 fragments, in-house library retrieval and literature research, 83 compounds were identified or tentatively characterized from Psoraleae Fructus, including 58 flavonoids, 11 coumarins, 4 terpenoid phenols, and 10 others. Sixteen of them were identified by comparison with reference compounds, and ten compounds may have not been reported from Psoraleae Fructus. This study achieved a rapid qualitative analysis on the chemical components in Psoraleae Fructus, which provided useful reference for elucidating its material basis and promoting the quality control.

Medicinal and edible plant Allium macrostemon Bunge for the treatment of testosterone-induced androgenetic alopecia in mice

Ethnopharmacological relevanceAllium macrostemon Bunge (AMB), a widely distributed wild garlic plant, possesses a variety of health-promoting properties. Androgenetic alopecia (AGA) is a common disorder that affects quality of life. Aim of the studyWe sought to investigate whether AMB stimulates hair regrowth in AGA mouse model, and clarify the underlying molecular mechanisms. Materials and methodsThe chemical constituents of AMB water extract were identified by ultra-high performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q/TOF-MS) analysis. Cell viability assay and Ki-67 immunostaining were undertaken to evaluate the impacts of AMB on human hair dermal papilla cell (HDPC) proliferation. Wound-healing assay was undertaken to assess cell migration. Flow cytometry and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay were performed to examine cell apoptosis. Western blotting, real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and immunostaining assays were undertaken to determine the impacts of AMB on Wnt/β-catenin signaling and growth factors expression in HDPC cells. AGA mouse model was induced by testosterone treatment. The effects of AMB on hair regeneration in AGA mice were demonstrated by hair growth measuring and histological scoring. The levels of β-catenin, p-GSK-3β, and Cyclin D1 in dorsal skin were measured. ResultsAMB promoted proliferation and migration, as well as the expression of growth factors in cultured HDPC cells. Meanwhile, AMB restrained apoptosis of HDPC cells by increasing the ratio of anti-apoptotic Bcl-2/pro-apoptotic Bax. Besides, AMB activated Wnt/β-catenin signaling and thereby enhancing growth factors expression as well as proliferation of HDPC cells, which was abolished by Wnt signaling inhibitor ICG-001. In addition, an increase of hair shaft elongation was observed in mice suffering from testosterone-induced AGA upon the treatment of AMB extract (1% and 3%). Consistent with the in vitro assays, AMB upregulated the Wnt/β-catenin signaling molecules in dorsal skin of AGA mice. ConclusionThis study demonstrated that AMB promoted HDPC cell proliferation and stimulated hair regrowth in AGA mice. Wnt/β-catenin signaling activation, which induced production of growth factors in hair follicles and, eventually, contributed to the influence of AMB on the hair regrowth. Our findings may contribute to effective utilization of AMB in alopecia treatment.

Comprehensive metabolic analyses provide new insights into primary and secondary metabolites in different tissues of Jianghua Kucha tea (Camellia sinensis var. assamica cv. Jianghua).

Jianghua Kucha (JHKC) is a special tea germplasm with enriched specialized secondary metabolites, including theacrine, non-epimeric flavanols and methylated flavanols. Moreover, primary metabolites provide precursors and energy for the production of secondary metabolites. However, the accumulation patterns of primary and secondary metabolites in different tissues of JHKC are unclear. The changes of primary and secondary metabolites and related metabolic pathways (primary and secondary metabolism) in different JHKC tissues (the bud, 1st-4th leaves, and new stem) were investigated via metabolomics analysis with ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS). Significant differences were observed in 68 primary and 51 secondary metabolites mainly related with the pathways of starch and sucrose, amino acids, caffeine, and flavanols metabolism and TCA cycle. The bud exhibited higher levels of glucose-6-phosphate, citric acid, most amino acids, theobromine, catechin-gallate, epicatechin-gallate, procyanidins, and theasinensins; the 1st leaf showed higher levels of caffeine and epigallocatechin-3-gallate; and the 4th leaf contained higher levels of most monosaccharides, theacrine, and epigallocatechin-3-O-(3"-O-methyl)-gallate. In addition, primary metabolites and important secondary metabolites had certain correlations. This study provides comprehensive insight into primary and secondary metabolites in JHKC and offers guidelines for efficiently utilizing specialized metabolites of JHKC in the future.

Unveiling the structure–activity relationships of ofloxacin degradation by Co3O4-activated peroxymonosulfate: From microstructures to exposed facets

As a fluoroquinolone antimicrobial agent, ofloxacin (OFX) has been widely used, consequently causing serious damage to the ecological environment. Advanced oxidation processes based on persulfate activation (SR-AOPs) are widely applied to degrade stubborn organic contaminants in wastewater treatment. Co3O4 is being extensively investigated as a promising persulfate catalyst in SR-AOPs. In this work, four Co3O4 polycrystals with specific morphologies were synthesized and used to activate peroxymonosulfate (PMS) for OFX degradation. The obtained samples and corresponding precursors were characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), and N2 adsorption/desorption isotherms. Experimental results showed that 30 mg·L-1 OFX could be completely removed in the lamella-like Co3O4/PMS system within 30 min under ordinary conditions. The underlying degradation mechanism was elaborated by high-resolution TEM (HRTEM) combined with density functional theory (DFT) calculations. Based on quenching experiments and the electron paramagnetic resonance (EPR) technique, sulfate radicals were identified as the dominant reactive oxygen species (ROS) in the lamella-like Co3O4/PMS system, and superoxide radicals, hydroxyl radicals, and singlet oxygen made minor contributions to the degradation of OFX. Possible degradation pathways were proposed based on DFT calculations and Fukui theory and confirmed by ultrahigh-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UHPLC-QTOF/MS). Furthermore, quantitative structure–activity relationship (QSAR) prediction was used to evaluate the developmental toxicity of the corresponding intermediates.

Integrating Metabolomics and Network Pharmacology to Explore the Mechanism of Tongmai Yangxin Pills in Ameliorating Doxorubicin-Induced Cardiotoxicity.

Doxorubicin (DOX) is a broad-spectrum chemotherapeutic drug used in clinical treatment of malignant tumors. It has a high anticancer activity but also high cardiotoxicity. The aim of this study was to explore the mechanism of Tongmai Yangxin pills (TMYXPs) in ameliorating DOX-induced cardiotoxicity through integrated metabolomics and network pharmacology. In this study, first, an ultrahigh-performance liquid chromatography-quadrupole-time-of-flight/mass spectrometry (UPLC-Q-TOF/MS) metabonomics strategy was established to obtain metabolite information and potential biomarkers were determined after data processing. Second, network pharmacological analysis was used to evaluate the active components, drug-disease targets, and key pathways of TMYXPs to alleviate DOX-induced cardiotoxicity. Targets from the network pharmacology analysis and metabolites from plasma metabolomics were jointly analyzed to select crucial metabolic pathways. Finally, the related proteins were verified by integrating the above results and the possible mechanism of TMYXPs to alleviate DOX-induced cardiotoxicity was studied. After metabolomics data processing, 17 different metabolites were screened, and it was found that TMYXPs played a role in myocardial protection mainly by affecting the tricarboxylic acid (TCA) cycle of myocardial cells. A total of 71 targets and 20 related pathways were screened out with network pharmacological analysis. Based on the combined analysis of 71 targets and different metabolites, TMYXPs probably played a role in myocardial protection through regulating upstream proteins of the insulin signaling pathway, MAPK signaling pathway, and p53 signaling pathway, as well as the regulation of metabolites related to energy metabolism. They then further affected the downstream Bax/Bcl-2-Cyt c-caspase-9 axis, inhibiting the myocardial cell apoptosis signaling pathway. The results of this study may contribute to the clinical application of TMYXPs in DOX-induced cardiotoxicity.

Determination of multiple mycotoxins in chili powder using cold-induced liquid–liquid extraction and Fe3O4@MWCNTs-NH2 coupled with UPLC-Q-TOF/MS

Food matrix interference is still a big challenge in analyzing multiclass mycotoxins. Herein, a novel cold-induced liquid–liquid extraction-magnetic solid phase extraction (CI-LLE-MSPE) coupled with ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS) method was explored for the simultaneous determination of multiple mycotoxins in chili powders. Fe3O4@MWCNTs-NH2 nanomaterials were prepared and characterized, and the factors affecting the MSPE process were investigated. Based on this, the CI-LLE-MSPE-UPLC-Q-TOF/MS method was established for determining ten mycotoxins in chili powders. The proffered technique eliminated the matrix interference effectively and demonstrated strong linearity (0.5–500 µg/kg, R2 ≥ 0.999), high sensitivity (limit of quantification was 0.5–1.5 µg/kg), and the recovery was 70.6%-111.7%. The extraction process is simpler than conventional methods, as the adsorbent can be separated using magnets, and reusable adsorbents are beneficial in reducing costs. In addition, the method can provide a valuable reference for pretreatment procedures for other complex matrices.

Dynamic variations in the chemical constituents of Tiebangchui stir-fried with Zanba by integrating UPLC-Q-TOF-MS based metabolomics and DESI-MSI

Tiebangchui, which mainly originates from the dried root of Aconitum pendulum Busch, is an important traditional Tibetan medicine and has long been used for the treatment of various types of cold and pain. However, the excellent pharmacological activities of Tiebangchui are accompanied by high toxicity caused by Aconitum alkaloids. To ensure clinical medication safety, Tiebangchui is only used after processing. Stir-frying with Zanba is an effective and unique processing method in traditional Tibetan medicine, but chemical variations in raw and processed Tiebangchui are rarely reported. In this study, an integrated method combining ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and desorption electrospray ionization mass spectrometry imaging (DESI-MSI) was applied to explore chemical constituent variations, transformation mechanisms, and in situ spatial distributions of metabolites in raw Tiebangchui and its series of processed products. Three important diterpenoid alkaloids were quantified by HPLC. Results revealed visible differences between raw Tiebangchui and its Zanba stir-fried products, and 60 min was demonstrated to be the proper processing time. A total of 64 components were detected, and 32 of them were considered as metabolic markers to distinguish raw Tiebangchui and its processed products stir-fried for different time. Chemical changes were also visualized by DESI-MSI: the contents of aconitine and 3-deoxyaconitine significantly decreased, and those of benzoylaconine, benzoyldeoxyaconine, 16-epi-pyrodeoxyaconitine, and 16-epi-pyroaconitine markedly increased. Transformations from diester-diterpenoid alkaloids to monoester-diterpenoid alkaloids through hydrolysis and pyrolysis were found to be the major detoxication processes during Zanba stir-frying. In summary, UPLC-Q-TOF-MS based metabolomics, DESI-MSI, and quantitative combined approach could be an efficient method to characterize chemical variations in Tiebangchui and its Zanba processed products with different stir-frying time. The wide application of this method would contribute to the process monitoring and safe control of Tiebangchui and other toxic Aconitum medicine.

Plasma and milk metabolomics in lactating sheep divergent for feed efficiency

Enhancing the ability of animals to convert feed into meat or milk by optimizing feed efficiency (FE) has become a priority in livestock research. Although untargeted metabolomics is increasingly used in this field and may improve our understanding of FE, no information in this regard is available in dairy ewes. This study was conducted to (1) discriminate sheep divergent for FE and (2) provide insights into the physiological mechanisms contributing to FE through high-throughput metabolomics. The ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q/TOF-MS) technique was applied to easily accessible animal fluids (plasma and milk) to assess whether their metabolome differs between high- and low-feed efficient lactating ewes (H-FE and L-FE groups, respectively; 8 animals/group). Blood and milk samples were collected on the last day of the 3-wk period used for FE estimation. A total of 793 features were detected in plasma and 334 in milk, with 100 and 38 of them, respectively, showing differences between H-FE and L-FE. The partial least-squares discriminant analysis separated both groups of animals regardless of the type of sample. Plasma allowed the detection of a greater number of differential features; however, results also supported the usefulness of milk, more easily accessible, to discriminate dairy sheep divergent for FE. Regarding pathway analysis, nitrogen metabolism (either anabolism or catabolism) seemed to play a central role in FE, with plasma and milk consistently indicating a great impact of AA metabolism. A potential influence of pathways related to energy/lipid metabolism on FE was also observed. The variable importance in the projection plot revealed 15 differential features in each matrix that contributed the most for the separation in H-FE and L-FE, such as l-proline and phosphatidylcholine 20:4e in plasma or l-pipecolic acid and phosphatidylethanolamine (18:2) in milk. Overall, untargeted metabolomics provided valuable information into metabolic pathways that may underlie FE in dairy ewes, with a special relevance of AA metabolism in determining this complex phenotype in the ovine. Further research is warranted to validate these findings.

Role of simulated in vitro gastrointestinal digestion on biotransformation and bioactivity of astragalosides from Radix Astragali

Radix Astragali (RA) is commonly used in Asian herbal therapy or food supply, and astragalosides and flavonoids are its major components with diverse pharmaceutical effects. To provide new information on the potential cardiovascular benefits of RA administered orally, the bioaccessibility of these compounds with relevant in vitro digestion parameters was determined for four digestion phases (oral, gastric, small and large intestines) by ultrahigh-performance liquid chromatography quadrupole time-of-flight-mass spectrometry (UPLC-Q-TOF/MS). Meanwhile, we compared the effects of digestion products on advanced glycation end products (AGEs)-induced intracellular reactive oxygen species (ROS) levels in a human arterial endothelial cells (HAECs) model, and studied the potential of RA against oxidative stress-related cardiovascular disease. The changes of saponins and flavonoids composition and antioxidant activity after digestion in intestines were mainly due to the astragaloside IV (AS-IV) biosynthesis involving saponins acetyl isomerization and deacetylation, and the flavonoid glycosides converted to aglycone by deglycosylation processes. All these results suggest that acetyl biotransformation of RA in small intestine directly influenced the response to oxidative stress, and might provide a reference for elucidation of the multi-component action after oral RA in cardiovascular health care.

The Effect of Epimedium Isopentenyl Flavonoids on the Broiler Gut Health Using Microbiomic and Metabolomic Analyses.

Epimedium (EM), also known as barrenwort, is a traditional medicinal plant rich in isopentenyl flavonols, which have beneficial biological activities and can improve human and animal health, but its mechanism is still unclear. In this study, ultra-high-performance liquid chromatography/quadrupole-time-of-flight-mass spectrometry (UHPLC-Q-TOF/MS) and ultra-high-performance liquid chromatography triple-quadrupole mass spectrometry (UHPLC-QqQ-MS/MS) were used to analyse the main components of EM, and isopentenyl flavonols such as Epimedin A, B, and C as well as Icariin were the major components of EM. Meanwhile, broilers were selected as model animals to illuminate the mechanism of Epimedium isopentenyl flavonols (EMIE) on gut health. The results showed that supplementation with 200 mg/kg EM improved the immune response, increased cecum short-chain fatty acids (SCFAs) and lactate concentrations, and improved nutrient digestibility in broilers. In addition, 16S rRNA sequencing showed that EMIE altered the composition of cecal microbiome, increasing the relative abundance of beneficial bacteria (Candidatus Soleaferrea and Lachbospiraceae NC2004 group and Butyricioccus) and reducing that of harmful bacteria (UBA1819, Negativibacillus, and Eisenbergiella). Metabolomic analysis identified 48 differential metabolites, of which Erosnin and Tyrosyl-Tryptophan were identified as core biomarkers. Erosnin and tyrosyl-tryptophan are potential biomarkers to evaluate the effects of EMIE. This shows that EMIE may regulate the cecum microbiota through Butyricicoccus, with changes in the relative abundance of the genera Eisenbergiella and Un. Peptostreptococcaceae affecting the serum metabolite levels of the host. EMIE is an excellent health product, and dietary isopentenyl flavonols, as bioactive components, can improve health by altering the microbiota structure and the plasma metabolite profiles. This study provides the scientific basis for the future application of EM in diets.

Profile of Human Milk Phospholipids at Different Lactation Stages with UPLC/Q-TOF-MS: Characterization, Distribution, and Differences.

Human milk phospholipids are important for the regular growth and development of infants. Ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS) was employed to qualitatively and quantitatively analyze 277 phospholipid molecular species in 112 human milk samples to obtain a detailed profile of human milk phospholipids along the lactation stage. MS/MS fragmentation patterns of sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and phosphatidylserine were characterized in detail. Phosphatidylcholine is the most dominant group, followed by sphingomyelin. PC(18:0/18:2), SM(d18:1/24:1), PE(18:0/18:0), PS(18:0/20:4), and PI(18:0/18:2) showed the highest average concentration among all of the phosphatidylcholine, sphingomyelin, phosphatidylethanolamine, phosphatidylserine, and phosphatidylinositol molecular species, respectively. The fatty acids attached to the phospholipid molecules were mainly palmitic, stearic, oleic, and linoleic acids, and the plasmalogens decreased along the lactation stage. The increase of sphingomyelins and phosphatidylethanolamines and the decrease of phosphatidylcholines are the key changes from colostrum to transitional milk; the increase of lysophosphatidylcholines and lysophosphatidylethanolamines and the continuous decrease of phosphatidylcholines are the vital changes from transitional milk to mature milk.

Lippia javanica leaf extract as an effective anti-corrosion agent against mild steel corrosion in 1 M HCl and its characterization by UHPLC/Q-TOF-MS spectroscopy and quantum chemical evaluation of its adsorption process on Fe(110)

A study was conducted to assess the anticorrosive potential of Lippia javanica leaf extract as a mild steel corrosion inhibitor in 1 M HCl acidic media. The chemical compositions of the acetone extract and its fractions were investigated by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOF-MS) with C29H36O15 (verbascoside) identified as the major constituent of L. javanica. The anticorrosive assessment included weight loss measurements as well as an examination of thermodynamic parameters. According to weight loss data, 800 ppm of L. javanica at 303 K achieved a maximum inhibition efficiency of 98.01%. Temperature studies revealed that, in the presence of the extract, inhibition efficiency decreased, and activation energies increased as inhibitor concentration increased. The ability of L. javanica to maintain an inhibition efficiency of around 90% even at elevated temperatures, as supported by immersion time studies, indicates its effectiveness against mild steel corrosion. Electrochemical studies, open circuit potential (OCP), potentiodynamic polarization (PDP), and electrochemical impedance spectroscopy (EIS) were conducted to study the inhibition mechanism of the extract. PDP results indicated that the cathodic polarization slope values were higher than anodic ones, indicating that the extract components act principally on the cathodic branch of the metal, suppressing the reduction of hydrogen ions. Impedance diagrams showed that increasing L. javanica concentration increased charge transfer resistance (Rct) and decreased double layer capacitance (Cdl). The morphology and the surface composition of the mild steel were examined by scanning electron microscopy with energy dispersive x-ray analysis (SEM/EDX) to verify the presence of inhibitor components. Water contact angle (WCA), UV–visible, and Fourier-transform infrared spectroscopy (FTIR) were carried out to establish the corrosion-inhibitive properties responsible for the inhibition capabilities of the plant extract. Adsorption of L. javanica components onto mild steel surfaces was modelled to acceptable parameters using the Langmuir adsorption isotherm for PDP, EIS, and weight loss data. In this work, quantum calculations were performed to study the adsorption of the main component on a clean Fe(110) surface using density functional theory (DFT) with the PBE exchange-correlation functional in the DMol3 program.

The effect of the purslane polyphenols on the structure of rabbit meat myofibrillar protein under malondialdehyde-induced oxidative stress.

The present research aimed to assess the protective effects of purslane polyphenols on rabbit meat myofibrillar protein (MP) under malondialdehyde (MDA)-induced oxidation. The major polyphenols in purslane extract were analyzed by ultrahigh performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS). To analyze the protective mechanism of purslane polyphenol monomers on MP in the oxidation system, we determined the retained MDA content in the suspension and analyzed the structure and nuclear magnetic resonance (NMR) of MP. The UPLC-QTOF-MS identification analysis indicated that purslane polyphenols mainly contained chlorogenic acid, caffeic acid, ferulic acid, rutin, kaempferol, and quercetin. Polyphenols in purslane can prevent MDA from binding to MP effectively. Chlorogenic acid showed the best inhibitory effect, and the retention content of MDA decreased to 54.79%, which was significantly lower than those of other groups. Purslane polyphenols had protective effects on sulfhydryl groups, free amino groups, intrinsic tryptophan fluorescence, α-helix structure, and morphology of MP. Moreover, the carbonyl contents, increase in surface hydrophobicity, and random coil contents of MP were reduced. In addition, the NMR results indicated that polyphenols inhibited the oxidation of protein induced by MDA through competitive inhibition. PRACTICAL APPLICATION: This study showed the efficiency of purslane polyphenol monomers in protecting the structure and function of myofibrillar protein under malondialdehyde-induced oxidative stress and their potential to preserve the nutritional value of foods rich in proteins and lipids.

Computational method for rapid screening of the metabolites of Pulsatilla chinensis in rats using UHPLC-Q-TOF/MS combined with mass spectrum-based orthogonal projection

Screening metabolites in vivo can be challenging due to the complexity of traditional Chinese medicine (TCM) and the ambiguous intracorporal process. To resolve this problem, we established the mass spectrum-based orthogonal projection (MSOP) method to differentiate prototype compounds from metabolites in vivo and applied it to the study of metabolites of Pulsatilla chinensis (PC). Initially, the validity and feasibility of the MSOP method were verified by using the ultra- high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) data of reference solution. Then, the MSOP method was applied to screen the metabolites of PC. A total of 63 metabolites were identified in vivo (urine, feces, bile, and plasma samples) and in vitro (intestinal bacteria biological sample). The results indicated that the main metabolic pathways of pentacyclic triterpenoids were demethylation, oxidation, dehydration, sulfation, and glucuronidation reactions. This study contributes to developing an integrated strategy based on chemometrics to characterize and classify the metabolism feature of pentacyclic triterpenoids of PC. This will support the scientific and rational application of PC in the clinic. The MSOP method based on the orthogonality of MS signals was used to differentiate the prototype compounds from metabolites in vivo. The method provides scientific and reliable support for fully understanding the metabolic fate of TCM.

Metabolomic profiles in serum and urine uncover novel biomarkers in children with nephrotic syndrome.

Nephrotic syndrome is common in children and adults worldwide, and steroid-sensitive nephrotic syndrome (SSNS) accounts for 80%. Aberrant metabolism involvement in early SSNS is sparsely studied, and its pathogenesis remains unclear. Therefore, the goal of this study was to investigate the changes in initiated SSNS patients-related metabolites through serum and urine metabolomics and discover the novel potential metabolites and metabolic pathways. Serum samples (27 SSNS and 56 controls) and urine samples (17 SSNS and 24 controls) were collected. Meanwhile, the non-targeted analyses were performed by ultra-high-performance liquid chromatography-quadrupole time of flight-mass spectrometry (UHPLC-QTOF-MS) to determine the changes in SSNS. We applied the causal inference model, the DoWhy model, to assess the causal effects of several selected metabolites. An ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to validate hits (D-mannitol, dulcitol, D-sorbitol, XMP, NADPH, NAD, bilirubin, and α-KG-like) in 41 SSNS and 43 controls. In addition, the metabolic pathways were explored. Compared to urine, the metabolism analysis of serum samples was more clearly discriminated at SSNS. 194 differential serum metabolites and five metabolic pathways were obtained in the SSNS group. Eight differential metabolites were identified by establishing the diagnostic model for SSNS, and four variables had a positive causal effect. After validation by targeted MS, except XMP, others have similar trends like the untargeted metabolic analysis. With untargeted metabolomics analysis and further targeted quantitative analysis, we found seven metabolites may be new biomarkers for risk prediction and early diagnosis for SSNS.