Quadrupole Orbitrap Mass Spectrometry Research Articles

A Novel Insight into Postmenopausal Hypercholesterolemia: Carnitine as a Key Player.

Background: Postmenopausal women (post-MW) are at a heightened risk of cardiovascular diseases, including hypercholesterolemia. This study aimed to investigate metabolomic variations to identify potential markers and targets for postmenopausal hypercholesterolemia. Methods: Sixty-two female volunteers aged 40-65 were recruited for this study. Metabolomic analysis using the Ultra-Performance Liquid Chromatography Quadrupole Orbitrap Mass Spectrometry (UPLC-Q-Orbitrap MS) platform was conducted to investigate changes in endogenous substances in premenopause (n = 25) and postmenopause (n = 37) women. Following ovariectomy surgery, menopausal mice were monitored for changes in their biomarker levels, and the integrity of the large artery walls in each treatment group was observed through hematoxylin and eosin staining. In vitro cellular models were utilized to assess variations in lipid metabolism, reactive oxygen species (ROS) levels, and changes in the levels of superoxide dismutase and glutathione peroxidase enzymes in different cell groups postintervention using Western blot analysis. Results: Treatment with carnitine in postmenopausal mouse models led to increased plasma cholesterol and carnitine levels, as well as indicators of arterial sclerosis. In HepG2 cells, carnitine treatment resulted in heightened lipid levels, elevated ROS production, and decreased antioxidant enzyme levels. Conclusions: The findings suggest that carnitine may serve as a potential risk marker or therapeutic target for postmenopausal hypercholesterolemia. This study provides valuable insights into cardiovascular conditions in post-MW and offers new avenues for therapeutic interventions. Continued research in this area is crucial to enhance our understanding of cardiovascular diseases in post-MW and to explore additional treatment options.

Metabolic Differences among Patients with Cirrhosis Using Q Exactive Hybrid Quadrupole Orbitrap Mass Spectrometry Technology.

The hospitalization and mortality rates of patients gradually increase following the onset and progression of liver cirrhosis (LC). We aimed to help define clinical stage and better target interventions by detecting the expression of specific metabolites in patients with different stages of LC via Q Exactive hybrid quadrupole orbitrap mass spectrometry (UPLC-Q-Exactive) technology. This noninterventional observation case-control study involved 139 patients with LC or acute-on-chronic liver failure (ACLF) in a Chinese hospital between October 2022 and April 2023. Serum specimens were analyzed for multiple metabolite levels using UPLC-Q-Exactive. Data were processed to screen for differentially accumulated metabolites (DAMs). Short time-series expression miner (STEM) analysis and enrichment analysis were performed to assess cirrhosis progression biomarkers. Following univariate and multivariate analyses, a Venn diagram indicated nine significant DAMs in common among groups. STEM analysis showed 8'-hydroxyabscisic acid, HDCA, pyruvate-3-phosphate, indospicine, eplerenone, and DEHP as significant; their levels first peaked [Child-Turcotte-Pugh (CTP) class B peaked] and then decreased with CTP grade aggravation. Significant differences among 8'-hydroxyabscisic acid, eplerenone, and DEHP were observed among LC comorbidities and between subgroups. Therefore, serum levels of six DAMs may characterize metabolomic changes, determine the severity of LC, and predict the development of ACLF.

Degradation of non-phthalate plasticisers in soils by liquid chromatography coupled to high resolution mass accuracy spectrometry: A comparison with sterile soil

The market for non-phthalate plasticisers (NPPs), such as adipates, citrates, phosphates, and carboxylates, has grown due to their low toxicity, but no studies have determined their metabolites during degradation processes in soil. This study evaluated the degradation of diethyl hexyl adipate (DEHA) and acetyl triethyl citrate (ATEC) in nonsterile soil, and the results were compared with those obtained from sterile soil, to determine whether microbial activity causes degradation in these NPPs. The analyses of these substances were performed using liquid chromatography coupled with quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap-HRMS). Plasticiser degradation followed a biphasic kinetic model, with a half-life time (DT50) of 9–12 days in non-sterile soil, but very slow in sterile soil (DT50 > 58 days). The degradation of target plasticisers in nonsterile soil resulted in the detection of eight metabolites of DEHA and ATEC, which were not detected in sterile soil. Among them, 4-[(2-ethylhexyl)oxy]-4-oxobutanoic acid (EHOBA), 4-((2-ethylhexyl)oxy)-4-oxo-1-carbonyl hexanoate (EHOCH), triethylcitrate (TEC) and N-acetyl glutamine (NAG) have been detected for the first time in this study. Most of these metabolites were detected after 14 days, increasing their concentration to 21 days, before completely degrading after 45 days. Nevertheless, mono–2-ethyl-5-hydroxyhexyl adipate (MEHHA), mono-2-ethyloxohexyl adipate (MEHOA), EHOCH, and diethyl citrate (DEC) were persistent in non-sterile soil (detected at 45 days) with an LD50 lower than their respective original molecules. Therefore, it is necessary to control the amount of these metabolites derived from the degradation of this type of plasticisers in the soil to avoid health risks, as they can end up in vegetables.

Rapid determination of 54 dye components in hair dyes by liquid chromatography coupled to quadrupole orbitrap high-resolution mass spectrometry.

Chemical hair dye components can have allergenic, reproductive, and carcinogenic risks. Detecting restricted and prohibited ingredients in these products is challenging due to product diversity, isomer separation, instability, and wide polarity range. A method was developed using HPLC-high-resolution mass spectrometry for the qualitative and quantitative analysis of 54 hair dye components in various products. Samples were extracted with a 70% methanol solution, ultrasonicated in an ice bath, centrifuged, filtered, diluted with 25% methanol solution and 25% methanol solution containing 0.05% D-isoascorbic acid. Separation was achieved using an ACE Excel 3 C18 column (2.1 mm × 150 mm, 3 μm) with analysis conducted via quadrupole Orbitrap mass spectrometry. It showed good linear correlations, with detection limits of 0.1-23.5 ng mL-1, and quantitation limits of 0.2-78.1 ng mL-1. Average recovery ranged from 60.0% to 118.4%, with repeatability from 4.0% to 14.9%. Stability was confirmed within 48 hours. When applied to 20 batches of commercially available hair dyes, 24 hair dye components were found within permissible levels. The method is crucial for quality control of hair dyes, covering 10 common prohibited and 44 permissible hair dye components outlined in the Safety and Technical Standards for Cosmetics (2015 Edition). Compared to the standard methods, it can separate isomers in a single mobile phase system within 15 minutes in positive ion mode while maintaining sensitivity for phenol, hydroquinone, and other components in negative ion mode. Moreover, the pre-treatment strategy significantly improved stability and accuracy, enabling precise analysis of the 54 hair dye components.

Zilpaterol in bovine liver, meat, heart, and kidney, determined by liquid chromatography–quadrupole-orbitrap high-resolution mass spectrometry

ABSTRACT An analytical method was developed for identification and quantification of zilpaterol in bovine liver, meat, heart, and kidney, using liquid chromatography coupled to quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap HRMS). It was validated in accordance with Commission Implementing Regulation (CIR) EU 2021/808 at six different concentrations, ranging from 0.1 to 5 μg/kg. The mean recoveries ranged from 71% to 99%, while the decision limit (CCα) and detection capability (CCβ) ranges were 0.11–0.12 μg/kg and 0.13–0.15 μg/kg, respectively. The method demonstrated good linearity (R2 > 0.9996) and the limits of detection (LODs) and of quantification (LOQs) were in the range of 0.015–0.061 μg/kg and 0.025–0.091 μg/kg, respectively. Out of 200 samples collected from local markets in Egypt, 17 contained zilpaterol residues. Liver samples revealed the highest detection frequency (26%), followed by meat (6%), at mean concentrations of 2.64 and 1.93 μg/kg, respectively.

Identification of chemical constituents and pharmacokinetic characteristics of Xiaoyan Tuire Granule in rats

Xiaoyan Tuire Granule is a type of Chinese patent medicine that has been proven effective in treating respiratory tract infections. However, while it has been successfully introduced into clinical use, more knowledge is still needed regarding its chemical components and pharmacokinetics. This study investigated the chemical profile in the medicine and rat plasma by ultra high-performance liquid chromatography coupled with Q Exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry (UHPLC-Orbitrap-MS/MS). Subsequently, it developed a validated ultra high-performance liquid chromatography coupled with quadrupole mass spectrometry (UHPLC-MS/MS) method for determining five components in rat plasma after oral administration of Xiaoyan Tuire Granule. As a result, a total of 106 constituents were inferred, including 9 terpenoids, 29 flavonoids, 33 organic acids, 12 phenylpropanoids and 23 other compounds. After administration, 86 compounds were inferred in rat plasma, including 73 prototypes and 13 metabolites. The metabolic pathways were primarily hydrogenation, glucuronic acid conjugation, sulfate conjugation, hydrolysis and methylation. The established method determined the contents of esculetin, esculin, isovitexin, caffeic acid and p-coumaric acid had a good separation, and all the legal verification met the requirements. The pharmacokinetic results indicate that the absorption rate of the five compounds in vivo was rapid, with a Tmax of less than 0.25 h, and the elimination rate was also fast, with a half-time (T1/2) ranging from 1.22 h to 2.19 h. It is worth noting that esculin and esculetin have similar half-time in vivo due to their structural similarities. Among these five compounds, the AUC0-∞ and MRT0-∞ of p-coumaric acid and esculetin were relatively higher, indicating higher exposure and longer residence time of both compounds in vivo. In conclusion, this paper researched the chemical constituents and pharmacokinetics of Xiaoyan Tuire Granule, which provided the reference for further study.

Optimisation and validation of liquid-liquid extraction of aminoglycosides from foods of animal origin

ABSTRACT This study focused on evaluating and optimising the liquid-liquid extraction (LLE) method for the multi-residue analysis of seven AGs in bovine liver, meat, milk, and kidney in conjunction with liquid chromatography coupled to a quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap HRMS) for quantification. Of all the hydrophilic interaction liquid chromatography (HILIC) stationary phases tested, the ZIC-HILIC column yielded the best performance. The method was validated according to Commission Implementing Regulation (CIR) EU 2021/808, which encompassed various concentration ranges from 25–7500 µg/kg. The mean recoveries ranged from 79% for liver matrix to 87% for meat matrix, with coefficients of variation (CV) below 18%. The method exhibited excellent linearity, as indicated by correlation coefficients (R2) greater than 0.9903 across all tested matrices. The decision limit (CCα) and detection capability (CCβ) varied from 62 to 5984 µg/kg and 71 to 6471 µg/kg, respectively. The limits of detection (LOD) and quantification (LOQ) were established between 1 and 45 µg/kg, and 2 and 75 µg/kg, respectively. No AG residues were detected in the 120 samples that were collected from different local markets in Egypt using the developed analytical method.

Comparative analysis of chemical elements and metabolites in diverse garlic varieties based on metabolomics and ionomics.

As a plant classified under the "medicine food homology" concept, garlic offers various health benefits and comes in many different varieties. In this study, the metabolite composition of different garlic varieties were analyzed using LC-MS/MS quadrupole-Orbitrap mass spectrometry and ICP-MS. A total of 30 chemical elements and 1256 metabolites were identified. Significant differences in chemical elements and metabolomics profiles were observed among the five garlic groups (VIP > 1.5). Compared to WG, PG contained 5 unique compounds, HG had 15 unique compounds, SCG had 18 unique compounds, and SBG had 26 unique compounds. Furthermore, the results showed that WG had smaller differences with PG and HG, but significant differences with SBG and SCG. KEGG analysis revealed metabolic pathways associated with the formation of differential metabolites. These findings uncover the differences and mechanisms in the composition of various garlic varieties, providing a theoretical foundation for distinguishing the nutritional components of different garlic types.

Fungicide Metabolite MS2 Spectral Libraries for Comprehensive Human Biomonitoring.

Fungicides undergo rapid metabolism and are excreted in the urine. There are few methods for screening these ubiquitous compounds, which have a high potential for human exposure. High-resolution mass spectrometry (HRMS) is a suitable technique to assess fungicide exposures; however, there is a lack of spectral libraries for fungicide annotation and in particular for downstream metabolites. We created spectral libraries for 32 fungicides for suspect screening. Fungicide standards were administered to mice, and 24-h urine was analyzed using hydrophilic interaction and reversed-phase chromatography coupled to hybrid quadrupole-orbitrap mass spectrometry. Suspect metabolite MS2 spectra for library creation were selected based on the ratio of exposed-to-control mouse urine. MS2 libraries were applied to urine collected from female university students (n = 73). Several tetraconazole and tebuconazole metabolites were detected in 3% (2/73) of the samples. The creation of comprehensive suspect screening MS2 libraries is a useful tool to detect fungicide exposure for human biomonitoring.

Degradation of Isotianil in Water and Soil: Kinetics, Degradation Pathways, Mechanisms, and Ecotoxicity Assessments.

Pesticides are transported and transformed in soil and can enter surface water through various pathways. They undergo hydrolysis, oxidation, and photoconversion in surface water. Isotianil is a new fungicide that effectively controls rice blast. However, there are limited reports on its degradation. Herein, the hydrolysis and photolysis of isotianil in water and its degradation in soil samples from five provinces of China were investigated. The degradation products of isotianil were identified using ultrahigh-performance liquid chromatography-Q exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry, and four compounds were discovered for the first time. The degradation pathways of isotianil were inferred, and the reaction active site and degradation mechanism of isotianil were clarified based on density functional theory calculations. The ecotoxicity of the degradation product M118 (aminobenzonitrile) was found to be moderate toward Daphnia magna, which was predicted and confirmed by Ecological Structure Activity Relationships and the experiment, respectively. The results of this study will contribute to a better understanding of the fate of isotianil in the environment.

In-vivo metabolic profiling of the Natural products Emodin and Emodin-8-O-β-D-glucoside in rats using liquid chromatography quadrupole Orbitrap mass spectrometry

Emodin (EM) and emodin-8-O-β-D-glucoside (EG) were found in various medicinal plants such as Rheum palmatum, Aloe vera, Polygonum multiflorum, and Polygonum cuspidatum. They have different pharmacological properties and are considered potentially toxic substances. It is necessary to identify the metabolites and their distribution in the body. In this study, a comprehensive analytical strategy was developed to characterize the metabolites of EM and EG in vivo using UHPLC-Q-Exactive Orbitrap MS. 190 metabolites were identified in the bio-samples, in addition to the commonly reported glycoside hydrolysis, hydrogenation, hydroxylation, glucuronide conjugation, and sulfation conjugation. We also discovered new pathways such as formylation, acetylation, glycol acylation, lactation, glycerolization, malonylation, glycerol acid acylation, hydroxyvalerylation, erythrosylation, glutaric acidfication, hydroxybenzoylation, glutamylation, hydroxyglutamylation, ascorbylation, aspartylglycylation, dihydroxyphenylglycylation, trihydroxyphenylglycylation, dihydroxymethoxyphenylglycylation, and ring-opening of EM. Cluster analysis revealed that each tissue in the EM and EG groups exhibited a high degree of similarity in their metabolic pathway preferences and bodily distributions, as they clustered together. Interestingly, the presence of glycol acylation, glycerolization, and glycerol acid acylation in the liver may be related to the lipid-lowering effects of EM and EG. These findings offer valuable insights for a more comprehensive understanding of the safety and efficacy of EM and EG, as well as valuable methods for metabolic characterization.

Integrated UHPLC-Q-exactive-orbitrap MS/MS and ABTS analyses screen antioxidant and anti‑counterfeiting marker of Eucommiae Cortex (Duzhong) for Pharmacopoeias

Eucommiae Cortex (Duzhong) has been collected in several Pharmacopoeias. However, its Pharmacopoeia quality-marker is outdated and unreasonable. Therefore, the study used database-aided ultra-high-performance liquid chromatography coupled with hybrid quadrupole-orbitrap tandem mass spectrometry (UHPLC-Q-Exactive-Orbitrap MS/MS) analysis to putatively identify 33 compounds from Eucommiae Cortex. Especially, 14 “unexcavated” compounds were detected in Eucommiae Cortex for the first time; Meanwhile, all monocaffeoylquinic acid isomers (i.e., 1-O-, 3-O-, 4-O-, and 5-O- caffeoylquinic acids) have been strictly distinguished from each other, and three dicaffeoylquinic acid isomers (1,3-O-, 3,5-O-, and 4,5-O- dicaffeoylquinic acids) have been distinguished as well. All identified compounds were further subjected to a semi-quantification analysis, through which cholesteryl acetate (4.11 ± 0.46 %), sucrose (3.53 ± 0.063 %), and (−)-syringaresinol (2.81 ± 1.31 %) were found to be highly abundant. Furthermore, the semi-quantification results were integrated with ABTS+• antioxidant assay to individually calculate the antioxidant contribution of all compounds. (−)-Syringaresinol, neochlorogenic acid, vanillic acid, chlorogenic acid, and (−) pinoresinol were proven to be the top five antioxidant contributors, and their contribution values were 97.50, 46.31, 40.57, 35.05, and 30.90. Of the top contributors, chlorogenic acid was proposed as an additional pharmacopoeia Q-marker for its good testability and traceability. The additional Q-marker not only enhances the pharmacological relevance, but also effectively recognizes Eucommiae Cortex from its counterfeiting via liquid chromatography analysis. Thus, these findings will help Pharmacopoeias to update its Q-markers.

Sanhan Huashi Formula and Its Bioactive Compounds Exert Antiviral and Anti-Inflammatory Effects on COVID-19

Sanhan Huashi formula (SHHS), a traditional Chinese medicine (TCM), has shown significant therapeutic effects on coronavirus disease 2019 (COVID-19) in clinical settings. However, its specific mechanism and components still require further clarification. In vitro experiments with Vero-E6 cells infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) demonstrated that SHHS effectively inhibited viral invasion and proliferation. Complementary in vivo experiments using K18-human angiotensin converting enzyme 2 (hACE2) mice exposed to virus-like particles (VLPs) further confirmed that SHHS impeded SARS-CoV-2 entry. Although SHHS did not demonstrate direct antiviral effects in K18-hACE2 mice challenged with SARS-CoV-2, it significantly alleviated pathological damage and decreased the expression of chemokines such as C–C motif ligand (CCL)-2, CCL-3, C–X–C motif ligand (CXCL)-1, CXCL-6, CXCL-9, CXCL-10, and CXCL-11 in the lungs, suggesting that SHHS exerts immunomodulatory and anti-inflammatory effects via the CCL-2–CXCL axis. Additional research using a lipopolysaccharide (LPS)-induced acute lung injury (ALI) and RAW264.7 cell model validated the ability of SHHS to reduce the levels of inflammatory biomarkers, including interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α). Using advanced analytical techniques such as ultrahigh-performance liquid chromatography coupled with linear trap quadrupole Orbitrap mass spectrometry (UHPLC-LTQ-Orbitrap-MS) and surface plasmon resonance (SPR), nodakenin was identified as a potent antiviral component of SHHS that targets the 3C-like protease (3CLpro), a finding supported by the hydrogen–deuterium exchange mass spectrometry (HDX-MS) and molecular docking analyses. Furthermore, nodakenin demonstrated a significant antiviral effect, reducing the viral load by more than 66%. This investigation reveals that SHHS can combat COVID-19 by inhibiting viral invasion and promoting anti-inflammatory effects.

Urine as a Biobased Fertilizer: The Netherlands as Case Study

AbstractHumankind has grown increasingly dependent on mineral fertilizer to feed a growing world population. However, nutrients are rarely recovered and recycled after consumption, leading to environmental waste issues now and potential supply issues in the future. Urine is rich in these nutrients and is therefore a prime candidate as a precursor for biobased fertilizers. This work presents a case study to remove potentially present contaminants from human urine using activated carbon adsorbents to adhere to Dutch fertilizer legislation. In the urine, three pharmaceuticals were identified by target analysis and 11 other contaminants by suspect screening using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF/MS). These were successfully removed by treatments with both granulated and powdered activated carbon, albeit with a loss of 16–17% and 2–4% of dissolved nutrients, respectively. Further screening of inorganic contaminants and persistent organic pollutants by inductively coupled plasma mass spectrometry (ICP-MS) and gas chromatography quadrupole-Orbitrap mass spectrometry (GC-Q-Orbitrap/MS) showed that all prerequisites for fertilizer status are met, paving the way for its future legal use as a biobased fertilizer in the Netherlands.

Metabolite Profiling Analysis of the Tongmai Sini Decoction in Rats after Oral Administration through UHPLC-Q-Exactive-MS/MS.

Tongmai Sini decoction (TSD), the classical prescriptions of traditional Chinese medicine, consisting of three commonly used herbal medicines, has been widely applied for the treatment of myocardial infarction and heart failure. However, the absorbed components and their metabolism in vivo of TSD still remain unknown. In this study, a reliable and effective method using ultra-performance liquid chromatography coupled with hybrid quadrupole-Orbitrap mass spectrometry (UHPLC-Q-Exactive-MS/MS) was employed to identify prototype components and metabolites in vivo (rat plasma and urine). Combined with mass defect filtering (MDF), dynamic background subtraction (DBS), and neutral loss filtering (NLF) data-mining tools, a total of thirty-two major compounds were selected and investigated for their metabolism in vivo. As a result, a total of 82 prototype compounds were identified or tentatively characterized in vivo, including 41 alkaloids, 35 phenolic compounds, 6 saponins. Meanwhile, A total of 65 metabolites (40 alkaloids and 25 phenolic compounds) were tentatively identified. The metabolic reactions were mainly hydrogenation, demethylation, hydroxylation, hydration, methylation, deoxylation, and sulfation. These findings will be beneficial for an in-depth understanding of the pharmacological mechanism and pharmacodynamic substance basis of TSD.

Quick and high-throughput quantification of β-agonist residues in bovine liver, meat, milk, kidney, poultry, and egg using dispersive solid phase extraction

A reliable liquid chromatography coupled to quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap HRMS) method was developed for the simultaneous identification and quantification of 13 β-agonist residues in bovine liver, meat, milk, kidney, poultry, and egg. Dispersive-solid phase extraction (d-SPE) using acetonitrile (ACN) was used to prepare the samples. The analyte in the extracts was separated on a reversed-phase Accucore aQ (50 mm × 2.1 mm, 2.6 μm) using a mobile phase of an aqueous solution containing 2 mM ammonium acetate and acetonitrile (ACN) 0.1 % formic acid. The method was validated in accordance with Commission Implementing Regulation (CIR) EU 2021/808 at six different concentrations ranging from 0.1 to 5 μg/kg. The mean recoveries ranged from 65 to 94 %, while repeatability and reproducibility values were all below 13 %. The linearity, as correlation coefficients (R2) ranged from 0.9955 to 0.9999. The decision limit (CCα) and detection capability (CCβ) ranges were 0.11–0.13 µg/kg and 0.12–0.15 µg/kg, respectively. The limits of detection (LOD) and limits of quantification (LOQ) were in the range of 0.004–0.048 μg/kg and 0.010–0.075 μg/kg, respectively. Of the 180 samples that were collected from local markets in Egypt, 21.11 % had β-agonist residues. The mean concentration (µg/kg) and detection frequency (%) of the most frequently found β-agonist in the samples were as follows: terbutaline (2.63 µg/kg and 90 %), ractopamine (5.14 µg/kg and 23.3 %). The method's applicability was verified by successfully completing two rounds of proficiency testing (PT).

Comparative study on chemical constituents of different medicinal parts of Lonicera japonica Thunb. Based on LC-MS combined with multivariate statistical analysis

Lonicerae japonicae flos (LJF), Lonicerae japonicae caulis (LJC), Lonicerae folium (LF) and Lonicerae fructus (LFR) are derived from Lonicera japonica Thunb., which are formed due to different medicinal parts. The efficacy of the 4 medicinal materials has similarities and differences. However, little attention has been paid to illustrate the differences in efficacy from the perspective of phytochemistry. In this study, ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry (UPLC-Q-Exactive-Orbitrap-MS) was used to qualitatively analyze the ingredients in 4 herbs. A total of 86 compounds were plausibly or unambiguously identified, there were 54 common components among the 4 medicinal materials, and each kind of medicinal materials had its own unique components. On the basis of qualitative analysis, ultra-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UPLC-QQQ-MS/MS) was used to quantitatively analyze 31 components contained in 4 medicinal materials, and principal component analysis (PCA), orthogonal partial least squares discriminant analysis (OPLS-DA) and other multivariate statistical analysis were furtherly performed for comparing the component contents. The results showed that the samples from the same parts were clustered into one group, and the samples from different medicinal parts were significantly different. The analysis of variable importance projection (VIP) value of the OPLS-DA model showed that 10 components including chlorogenic acid, secologanic acid, isochlorogenic acid A, loganin, lonicerin, loganic acid, secoxyloganin, sweroside, luteolin and rhoifolin were the main difference components among the 4 medicinal materials. The study not only lays a solid foundation for the intrinsic quality control of 4 medicinal materials and the study of different effects of the 4 medicinal materials at the phytochemical level, but also provides a basis for more rational utilization of various parts of L. japonica and expansion of medicinal resources.

Exploring the gut microbiota mediated biotransformation of Senecio scandens Buch.-Ham.: Insights from metabolite spectrum with UHPLC-Q-Orbitrap HRMS and bioinformatics analysis of gut microbiota metabolites

Senecio scandens Buch.-Ham., a traditional Chinese medicine commonly used clinically, exhibits various pharmacological properties, including anti-inflammatory, anti-tumor, antiviral, and antibacterial activities. However, its water extracts' chemical components and metabolites are inadequately understood, limiting further research. In this study, the chemical components and metabolism processes of Senecio scandens, both in vivo (plasma, feces, urine, and bile) and in vitro (gut microbiota and liver microsomes), were characterized based on ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap high-resolution mass spectrometry. Additionally, metabolites detectable in fecal samples and intestinal microbiota incubated but absent in liver microsomes were identified as characteristic metabolites of intestinal microbiota. The targets of the characteristic metabolites of intestinal microbiota were collected, followed by exploration of potential pathways through KEGG enrichment analysis. As a result, a total of 133 chemical components were preliminarily identified, including 35 organic acids, 21 alkaloids, 19 flavonoids and their glycosides, 17 phenylpropanoids, 10 jacaranda ketones, and 31 other compounds. Notably, 12 of these were potentially novel compounds. In addition, 39 prototype components in rats and 109 metabolites were identified and characterized, including 102 in vivo and 52 metabolites in vitro (51 in rat gut microbiota and 24 in rat liver microsomes). The main metabolic pathways include oxidation, reduction, hydrolysis, methylation, glucuronidation, sulfonation, and acetylation reactions. Furthermore, KEGG enrichment analysis revealed that the characteristic metabolites of intestinal microbiota may be related to the ErbB, FoxO, mTOR, and MAPK signaling pathways, exhibiting anti-inflammatory and anti-tumor effects. In summary, the chemical components and metabolites of Senecio scandens were comprehensively identified using a rapid and accurate method, providing a scientific basis for the in-depth study of the material basis and its clinical application of Senecio scandens.

Multiclass method for detecting 41 antibiotic residues in bovine liver, muscle, and milk using LC-Q-Orbitrap-HRMS

This study developed and validated an analytical method for the simultaneous identification and quantification of 41 veterinary antibiotic drugs from 9 different classes, including sulfonamides, quinolones, tetracyclines, penicillins, macrolides, nitroimidazoles, cephalosporins, diaminopyrimidines, and amphenicol, in animal-derived food products. The method used modified QuEChERS and liquid chromatography coupled to quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap HRMS). The method was validated in accordance with Commission Implementing Regulation (CIR) EU 2021/808 at five different concentrations ranging from 0.075 to 10,000 μg kg−1. The mean recoveries ranged from 58 % to 123 %, while repeatability values were all below 21 %. The calibration curves showed good linearity, with correlation coefficients (R2) ranging from 0.9905 to 0.9999. The limits of detection (LOD) and limits of quantification (LOQ) were in the range of 0.005–43.0 μg kg−1 and 0.008–71.7 μg kg−1, respectively. The decision limit (CCα) and detection capability (CCβ) ranges were 0.37–2347 µg/kg and 0.44–3099 µg/kg, respectively. Out of the 1330 samples collected from Egyptian local markets, 1.20 % had antibiotic residues. The applicability of the developed method was proven through successful four proficiency testing (PT). The proposed method was demonstrated to be reliable for the simultaneous analysis of multiclass veterinary drugs in bovine liver, muscle tissue, and milk.

Optimization and validation of metabolomics methods for feline urine and serum towards application in veterinary medicine

BackgroundMetabolomics is an emerging and powerful technology that offers a comprehensive view of an organism's physiological status. Although widely applied in human medicine, it is only recently making its introduction in veterinary medicine. As a result, validated metabolomics protocols in feline medicine are lacking at the moment. Since biological interpretation of metabolomics data can be misled by the extraction method used, species and matrix-specific optimized and validated metabolomic protocols are sorely needed. ResultsSystematic optimization was performed using fractional factorial experiments for both serum (n = 57) and urine (n = 24), evaluating dilution for both matrices, and aliquot and solvent volume, protein precipitation time and temperature for serum. For the targeted (n = 76) and untargeted (n = 1949) validation of serum respectively, excellent instrumental, intra-assay and inter-day precision were observed (CV ≤ 15% or 30%, respectively). Linearity deemed sufficient both targeted and untargeted (R2 ≥ 0.99 or 0.90, respectively). An appropriate targeted recovery between 70 and 130% was achieved. For the targeted (n = 69) and untargeted (n = 2348) validation of the urinary protocol, excellent instrumental and intra-assay precision were obtained (CV ≤ 15% or 30%, respectively). Subsequently, the discriminative ability of our metabolomics methods was confirmed for feline chronic kidney disease (CKD) by univariate statistics (n = 41 significant metabolites for serum, and n = 55 for urine, p-value<0.05) and validated OPLS-DA models (R2(Y) > 0.95, Q2(Y) > 0.65, p-value<0.001 for both matrices). SignificanceThis study is the first to present an optimized and validated wholistic metabolomics methods for feline serum and urine using ultra-high performance liquid chromatography coupled to quadrupole-Orbitrap high-resolution mass spectrometry. This robust methodology opens avenues for biomarker panel selection and a deeper understanding of feline CKD pathophysiology and other feline applications.