Q-Exactive Orbitrap Mass Spectrometry Research Articles

Qualitative analysis of licorice and strychnine decoction before and after combination using UPLC-QE-Orbitrap-MS.

Glycyrrhiza glabra L. (GG) and Strychnos nux-vomica L. (NV) are traditional Chinese medicines (TCMs). Changes in the chemical composition may occur before and after the GG-NV compatibility. Ultra-performance liquid chromatography Q-exactive Orbitrap mass spectrometry (UPLC-QE-Orbitrap-MS) was applied here to study the difference in the components of the GG and NV decoctions before and after they were combined. The changes in the chemical composition of GG and NV before and after the combination were determined. The precise molecular weight, retention time, and fragment ion peak of the different components of the decoctions before and after compatibility were obtained through UPLC-QE-Orbitrap-MS. Differential analysis methods, such as principal component analysis, were used for comparison. In the positive ion mode, 200 new components were added, whereas six components were lost. In the negative ion mode, 144 new compounds were identified, whereas three components were missing. The compatibility difference between GG and NV was studied through UPLC-QE-Orbitrap-MS. The chemical composition of GG and NV changed before and after compatibility, and a class of compounds different from GG and NV was identified in the co-decoction. This study provides an experimental basis for subsequent research into detoxification mechanisms of the GG-NV combination and offers a new analytical method for investigating the compatibility of various other TCM pairs.

Aerosol inhalation of total ginsenosides repairs acute lung injury and inhibits pulmonary fibrosis through SMAD2 signaling-mediated mechanism

BackgroundPulmonary fibrosis (PF) is a progressive lung disease caused by previous acute lung injury (ALI), but there is currently no satisfactory therapy available. Aerosol inhalation of medicine is an effective way for treating PF. Total ginsenosides (TG) shows potential for the treatment of ALI and PF, but the effects of inhaled TG remain unclear. PurposeTo determine the therapeutic effects of TG in ALI and PF, to assess the superiority of the inhaled form of TG over the routine form, and to clarify the mechanism of action of inhaled TG. MethodsUltrahigh-performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry (UPLC-QE-MS) was applied to determine the chemoprofile of TG. A mouse model of ALI and PF was established to evaluate the effects of inhaled TG by using bronchoalveolar lavage fluid (BALF) analysis, histopathological observation, hydroxyproline assay, and immunohistochemical analysis. Primary mouse lung fibroblasts (MLF) and human lung fibroblast cell line (HFL1) were applied to determine the in vitro effects and mechanism of TG by using cell viability assay, quantitative real time PCR (qPCR) assay, and western blot (WB) analysis. ResultsThe UPLC-QE-MS results revealed the main types of ginsenosides in TG, including Re (14.15 ± 0.42%), Rd (8.42 ± 0.49%), Rg1 (6.22 ± 0.42%), Rb3 (3.28 ± 0.01%), Rb2 (3.09 ± 0.00%), Rc (2.33 ± 0.01%), Rg2 (2.09 ± 0.04%), Rb1 (1.43 ± 0.24%), and Rf (0.13 ± 0.06%). Inhaled TG, at dosages of 10, 20, and 30 mg/kg significantly alleviated both ALI and PF in mice. Analyses of BALF and HE staining revealed that TG modulated the levels of IFN-γ, IL-1β, and TGF-β1, reduced inflammatory cell infiltration, and restored the alveolar architecture of the lung tissues. Furthermore, HE and Masson's trichrome staining demonstrated that TG markedly decreased fibroblastic foci and collagen fiber deposition, evidenced by the reduction of blue-stained collagen fibers. Hydroxyproline assay and immunohistochemical analyses indicated that TG significantly decreased hydroxyproline level and down-regulated the expression of Col1a1, Col3a1, and α-sma. The inhaled administration of TG demonstrated enhanced efficacy over the oral route when comparable doses were used. Additionally, inhaled TG showed superior safety and therapeutic profiles compared to pirfenidone, as evidenced by a CCK8 assay, which confirmed that TG concentrations ranging from 20 to 120 μg/ml were non-cytotoxic. qPCR and WB analyses revealed that TG, at concentrations of 25, 50, and 100 μg/ml, significantly suppressed the phosphorylation of smad2 induced by TGF-β1 and down-regulated the expression of fibrotic genes and proteins, including α-sma, Col1a1, Col3a1, and FN1, suggesting an anti-fibrotic mechanism mediated by the smad2 signaling pathway. In vitro, TG's safety and efficacy were also found to be superior to those of pirfenidone. ConclusionsThis study demonstrates, for the first time, the therapeutic efficacy of inhaled TG in treating ALI and PF. Inhaled TG effectively inhibits inflammation and reduces collagen deposition, with a particular emphasis on its role in modulating the Smad2 signaling pathway, which is implicated in the anti-fibrotic mechanism of TG. The study also highlights the superiority of inhaled TG over the oral route and its favorable safety profile in comparison to pirfenidone, positioning it as an ideal alternative for ALI and PF therapy.

A comprehensive study of Chuanwang Xiaoyan capsule based on characterization of chemical constituents in vitro and in vivo using ultra-high-performance liquid chromatography-quadrupole-Exactive Orbitrap mass spectrometry and pharmacokinetic study of multiple components in rats using ultra-high-performance liquid chromatography-triple quadrupole-tandem mass spectrometry.

Chuanwang Xiaoyan (CWXY) capsule is primarily used to treat a variety of acute and chronic inflammations, including acute and chronic pharyngitis and tonsillitis. However, a systematic study of its chemical constituents is still not available. This study evaluated the chemical constituents in vitro and metabolite profiles in vivo of CWXY using ultra-high-performance liquid chromatography (UHPLC) coupled with Q-Exactive Orbitrap mass spectrometry, and the pharmacokinetic behaviors of the nine main components in rats were detected using ultra-high-performance liquid chromatography-triple quadrupole-mass spectrometry (UPLC-QQQ-MS/MS). A total of 92 chemical constituents in CWXY were preliminarily identified in vitro. After oral administration to rats, 56 prototype components and 128 metabolites of CWXY were detected in the biological samples of rat plasma, urine, bile, and feces. Of these prototype components and metabolites, seven new compounds, namely M15, M16, M25, M30, M31, M71, and M128, were detected for the first time. The quantitation method of nine components in rat plasma was developed using a pharmacokinetic study. To the best of our knowledge, this study was the first to investigate the pharmacokinetic behavior of triumbelletin.

Impact of extraction method on the lipids of Himalayan marmot oil with ultrahigh-performance liquid chromatography Q-Exactive Orbitrap mass spectrometry analysis.

Himalayan marmot oil (SPO) has been used for pharmaceutical purposes for centuries, but its composition is still unclear. The bioactivity of SPO highly depends on the techniques used for its processing. This study focused on the comprehensive lipidomics of SPO, especially on the ones derived from dry rendering, wet rendering, cold pressing, and ultrasound-assisted solvent extraction. We performed lipid profiling of SPO acquired by different extraction methods using ultrahigh-performance liquid chromatography Q-Exactive Orbitrap mass spectrometry, and 17 classes of lipids (2 BMPs, 12 LysoPCs, 9 LysoPEs, 41 PCs, 24 PEs, 23 Plasmenyl-PCs, 10 Plasmenyl-PEs, 10 MGs, 63 DGs, 187 TGs, 2 MGDGs, 3 Cer[NDS]s, 22 Cer[NS]s, 2 GlcCer[NS]s, 14 SMs, 14 CEs, and 6 AcylCarnitines) were characterized. Fifty-five lipids were differentially altered (VIP > 1.5, p < 0.05) between the extraction techniques, which can be used as potential biomarkers to differentiate SPO extracted by various methods. Additionally, the contents of oleic acid and arachidic acid were abundant in all samples that may suggest their medicinal values and are conducive to in-depth research. These findings reveal the alterations of lipid profile and free fatty acid composition in SPO obtained with different extraction methods, providing a theoretical foundation for investigating its important components as functional factors in medicines and cosmetics.

Insight into pyrrolizidine alkaloids degradation and the chemical structures of their degradation products using ultra high performance liquid chromatography and Q-Exactive Orbitrap mass spectrometry

Pyrrolizidine alkaloids (PAs), released into the environment by donor plants, are absorbed by crops or transported by animals, posing a global food safety concern. Photolysis is an effective way to eliminate harmful substances in the environment or food. Photolysis happens as PAs move among plants, environment and crops. In this study, we first investigated the photolysis and hydrolysis of 15 PAs and identified their degradation products via ultra-high performance liquid chromatography and Q-Exactive Orbitrap mass spectrometry. PAs were degraded under UV radiation but minimally affected by visible light from a xenon lamp, and solvent pH had little impact on their photolysis. PAs were stable in neutral and acidic solutions but degraded by 50% within 24 h in alkaline conditions. The degradation products of PAs were mainly PAs/PANOs isomers and some minor byproducts. Cytotoxicity and computational analysis revealed isomers had similar toxicity, with minor products being less toxic. This study is a precursor for revealing the potential PAs degradation dynamics in the environment and food products, providing a reference for systematic evaluations of potential health and ecological risks of their degradation products.

UHPLC Q-Exactive Orbitrap MS based metabolomics and biological activities of Camellia fascicularis from different geographical regions

Camellia fascicularis is a specific botanical resource from Yunnan province, China. It is rich in several active ingredients and nutrients and has both ornamental and medicinal contexts. However, different geographical environments affect the accumulation of metabolites, leading to quality differences. In this study, the metabolites of C. fascicularis from three geographical regions, namely Maguan (MG), Hekou (HK), and Gejiu (GJ), were investigated using ultra-high-performance liquid chromatography Q-Exactive Orbitrap mass spectrometry (UHPLC Q-Exactive Orbitrap MS) and the antioxidant and antitumor capacities were thoroughly assessed. The metabolomics analysis showed that 232, 215, and 83 significantly differential metabolites (SDMs) of three comparative groups, MG vs. HK, GJ vs. MG, and GJ vs. HK, had been screened in POS, respectively, whereas 29, 22, and 11 had been screened in NEG, respectively, with flavonoids as the main active SDMs. In vitro antioxidant and antitumor activities for MG, HK, and GJ were different, and HK exhibited the strongest antitumor ability. The correlation analysis indicated that the differences in activities of C. fascicularis from different geographical regions associated with flavonoids, such as isoquercitrin, hesperetin 7-glucoside, cosmosiin, mulberrin, vicenin 2, and corymboside in SDMs. This research provides a basis for evaluating germplasm resources and exploring the potential for development derived from the medicinal plant C. fascicularis.

Screening and Characterization of an α-Amylase Inhibitor from Carya cathayensis Sarg. Peel.

Inhibiting α-amylase can lower postprandial blood glucose levels and delay glucose absorption, offering an effective approach for the development of antidiabetic diets. In this study, an active constituent with inhibitory activity against α-amylase was isolated and purified by bioassay-guided fractionation from Carya cathayensis Sarg. peel (CCSP). The active constituent was identified by NMR and Q-Exactive Orbitrap Mass Spectrometry as 5-O-p-coumaroylquinic acid (5-CQA). 5-CQA possessed strong inhibitory activity against α-amylase, with an IC50 value of 69.39 µM. In addition, the results of the kinetic study indicated that 5-CQA was a potent, reversible, noncompetitive inhibitor against α-amylase. The findings indicate that 5-CQA derived from CCSP has potential as a novel inhibitor against α-amylase, which can help mitigate postprandial blood sugar spikes, making it suitable for inclusion in antidiabetic diets.

Chlorinated paraffins (CPs) in vegetable oils from the Dutch market and the effects of the refining process on their levels

Chlorinated paraffins (CPs) are of growing public health concern because of their ubiquitous presence in food samples. To get more insight into levels of CPs in vegetable oils from the Dutch market and human exposure via these oils, two series of samples were analysed in this study for the presence of CPs by UHPLC coupled with Q-Exactive Orbitrap mass spectrometry: 1) 39 vegetable oils from local supermarkets; 2) 13 paired crude and refined vegetable oils obtained from a contract refining company. Short chain CPs (SCCPs) and medium chain CPs (MCCPs) were detected in vegetable oils from supermarkets with an overall range from <LOQ to 78 ng/g lipid weight (lw) and <LOQ to 391 ng/g lw, respectively. The tridecane and tetradecane groups dominated in SCCPs and MCCPs, respectively. As regards to paired crude and refined oils from a refining company, a marked decrease in concentrations of both SCCPs and MCCPs were detected by the refining process (average reduction: 82% and 69% for SCCPs and MCCPs, respectively). Based on the data obtained, it can be concluded that CPs with a relatively short carbon chain length and low chlorination degree are more likely to be removed in the refining process. The estimated margins of exposure (MoEs) due to oil consumption alone were all higher than 1000, indicating no health concern from CPs for the Dutch population.

Screening the components in multi-biological samples and the comparative pharmacokinetic study in healthy and depression model rats of Suan-Zao-Ren decoction combined with a network pharmacology

Ethnopharmacological relevanceSuanzaoren Decoction (SZRD) is a classic traditional Chinese prescription, which has been commonly used for treating insomnia, depression and other nerve system diseases for a long time. Aim of this studyThe present study aimed to explore the metabolic profiles in multi-biological samples and pharmacokinetic mechanism between healthy and depression model rats combined with a network pharmacology approach after administration of SZRD. Materials and methodsIn our study, an ultra-high performance liquid chromatography (UPLC)-Q-Exactive Orbitrap Mass Spectrometry method was firstly used to study the prototype components and metabolites of SZRD in plasma, brain, urine, and feces between healthy and depressed rats. The possible metabolic pathways were also speculated. Then a network pharmacological study was conducted on the components in the plasma of model rats. According to the above components screened by network pharmacology and the other reported representative active components, the comparative pharmacokinetic study was established for the simultaneous determination of mangiferin, spinosin, ferulic acid, liquiritin, formononetin. magnoflorine and isoliquiritin between healthy and depression model rats. Finally, molecular docking was used to validate the binding affinity between key potential targets and active components in pharmacokinetics. ResultsA total of 115 components were identified in healthy rats, and 101 components were identified in model rats. The prototype components and metabolites in plasma, brain, urine, and feces were also distinguished. The main metabolic pathways included phase I and phase II metabolic reactions, such as dehydrogenation, oxidation, hydroxylation, gluconaldehyde conjugation, glutathione conjugation and so on. These results provided a basis for the further study of antidepressive pharmacokinetic and pharmacological action in SZRD. Then, according to the degree value of network pharmacological study, it was predicted that 10 components and 10 core targets, which involved in the critical pathways such as neuroactive ligand-receptor interaction, cyclic adenosine monophosphate (cAMP) signaling pathway, serotonergic synapse, phosphatidylinositol-3 kinase (PI3K)-Akt signaling pathway, etc. Finally, the established pharmacokinetic method was successfully applied to compare the pharmacokinetic behavior of these 7 active components in plasma of healthy and depressed rats after oral administration of SZRD. It showed that except magnoflorine, the pharmacokinetic parameters of each component were different between healthy and depressed rats. Molecular docking analysis also indicated that the active compounds in pharmacokinetics could bind tightly to the key targets of network pharmacological study. ConclusionThis study may provide important information for studying the action mechanism of SZRD in treating depression.

Detection and quantification of the metabolite Ac-Tβ1-14 in in vitro experiments and urine of rats treated with Ac-Tβ4: A potential biomarker of Ac-Tβ4 for doping tests.

Thymosin β4 (Tβ4) was reported to exert various beneficial bioactivities such as tissue repair, anti-inflammation, and reduced scar formation, and it is listed on the prohibited substances in sports by the World Anti-Doping Agency. However, no metabolism studies of Tβ4 were reported yet. Previously, our lab reported in in vitro experiment that a total of 13 metabolites were found by using multiple enzymes, and six metabolites (Ac-Tβ31-43 , Ac-Tβ17-43 , Ac-Tβ1-11 , Ac-Tβ1-14 , Ac-Tβ1-15 , and Ac-Tβ1-17 ) were confirmed by comparing with the synthetic standards. This study was aimed at identifying new metabolites of Tβ4 leucine aminopeptidase (LAP), human kidney microsomes (HKM), cultured huvec cells, and rats after administration of Tβ4 protein to develop biomarkers for detecting doping drugs in sports. A method for detecting and quantifying Ac-Tβ1-14 was developed and validated using Q-Exactive orbitrap mass spectrometry. The limit of detection (LOD) and limit of quantification (LOQ) of the Ac-Tβ1-14 were 0.19 and 0.58 ng/mL, respectively, and showed a good linearity (r2 = 0.9998). As a result, among the six metabolites above, Ac-Tβ1-14 , as a common metabolite, was found in LAP, HKM, huvec cells exposed to Tβ4, and the urine of rats intraperitoneally treated with 20-mg/kg Tβ4. And the metabolite Ac-Tβ1-14 was quantitatively determined by 48 h in rats, with the highest concentration occurring between 0 and 6h. Ac-Tβ1-14 was not detected in non-treated control groups, including human blank urine. These results suggest that Ac-Tβ1-14 in urine is a potential biomarker for screening the parent Tβ4 in doping tests.

Alterations in Faecal and Serum Metabolic Profiles in Patients with Neovascular Age-Related Macular Degeneration

Neovascular age-related macular degeneration (nAMD) is a common and multifactorial disease in the elderly that may lead to irreversible vision loss; yet the pathogenesis of AMD remains unclear. In this study, nontargeted metabolomics profiling using ultra-performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry was applied to discover the metabolic feature differences in both faeces and serum samples between Chinese nonobese subjects with and without nAMD. In faecal samples, a total of 18 metabolites were significantly altered in nAMD patients, and metabolic dysregulations were prominently involved in glycerolipid metabolism and nicotinate and nicotinamide metabolism. In serum samples, a total of 29 differential metabolites were founded, involved in caffeine metabolism, biosynthesis of unsaturated fatty acids, and purine metabolism. Two faecal metabolites (palmitoyl ethanolamide and uridine) and three serum metabolites (4-hydroxybenzoic acid, adrenic acid, and palmitic acid) were selected as potential biomarkers for nAMD. Additionally, the significant correlations among dysregulated neuroprotective, antineuroinflammatory, or fatty acid metabolites in faecal and serum and IM dysbiosis were found. This comprehensive metabolomics study of faeces and serum samples showed that alterations in IM-mediated neuroprotective metabolites may be involved in the pathophysiology of AMD, offering IM-based nutritional therapeutic targets for nAMD.

Qualitative analysis on the transdermal absorption from the dichloromethane extract of the Sambucus adnata wall. based on UPLC-Q-Exactive Orbitrap/MS

Sambucus adnata Wall.(SAW) has been used to treat osteoarthritis by the Yi nationality in China. The present study established an overall identification strategy based on ultra-high performance liquid chromatography-tandem Q-Exactive Orbitrap mass spectrometry (UPLC-Q-Exactive Orbitrap/MS) method to characterize the multiple chemical constituents of SAW before and after percutaneous penetration. Nineteen compounds, including triterpenoids, fatty acids, lignans, flavonoid, and amide, were tentatively identified in the dichloromethane extract of SAW, while fourteen ingredients penetrated the skin. Among them, eleven components were reported for the first time in SAW.

Equivalence of combined decoction and mixed single decoctions of Gegen Qinlian Decoction in alleviating chemotherapy-associated diarrhea

This study compared the chemical profiles, component content, dry paste yield, and pharmacological effects of samples obtained from the mixed single decoctions and the combined decoction of Gegen Qinlian Decoction(GQD), aiming to provide an experimental foundation for evaluating the equivalence of the two decocting methods and the suitability of TCM formula granules in clinical application. The same decoction process was used to prepare the combined decoction and mixed single decoctions of GQD. Ultra-performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was employed to compare the chemical profiles between the two groups. High-performance liquid chromatography(HPLC) was used to compare the content of nine characteristic components between the two groups. Then, a delayed diarrhea mouse model induced by irinotecan was established to compare the pharmacological effects of the two groups on chemotherapy-induced diarrhea. The UPLC-Q-Exactive Orbitrap MS in ESI~+ and ESI~- modes identified 59 chemical components in the compound decoction and mixed single decoctions, which showed no obvious differences in component species. The content of baicalin and wogonoside was higher in the compound decoction, while that of puerarin, daidzein-8-C-apiosylglucoside, berberine, epiberberine, wogonin, glycyrrhizic acid, and daidzein was higher in the mixed single decoctions. Further statistical analysis revealed no significant difference in the content of the nine characteristic components between the compound decoction and the mixed single decoctions. The dry paste yield had no significant difference between the two groups. Compared with the model group, both compound decoction and mixed single decoctions alleviated the weight loss and reduced diarrhea index in mice. Both of them lowered the levels of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), cyclooxygenase-2(COX-2), intercellular adhesion molecule-1(ICAM-1), interleukin-10(IL-10), malondialdehyde(MDA), and nitric oxide(NO) in the colon tissue. Furthermore, they significantly increased the levels of glutathione peroxidase(GSH-Px) and superoxide dismutase(SOD). Hematoxylin-eosin(HE) staining showed that colon tissue cells were tightly arranged with clear nuclei in both groups without obvious difference. The compound decoction and mixed single decoctions showed no significant differences in chemical component species, content of nine characteristic components, dry paste yield, or the pharmacological effects on alleviating chemotherapy-induced diarrhea. The findings provide a reference for evaluating the flexibility and superiority of combined or single decocting method in the preparation of TCM decoctions or formula granules.

Determination of subpicogram levels of airborne polycyclic aromatic hydrocarbons for personal exposure monitoring assessment

A method based on the use of GC coupled to Q-exactive Orbitrap mass spectrometry (GC-Orbitrap-MS) has been developed for the analysis of polycyclic aromatic hydrocarbons (PAHs) at sub-picogram levels. Outdoor ambient air particulate matter (PM2.5) and standard reference materials (SRM2260a) were analyzed in full scan mode showing low instrumental uncertainties (1–22%) and high linearity over a wide concentration range (0.5 pg and 500 pg/μL). Good reproducibility was obtained compared to the use of conventional single quadrupole GC–MS of PM samples. The quantification limit of the GC-Orbitrap-MS method for full scan analysis of PAHs in outdoor ambient air PM samples was 0.5 pg/μL. This low limit allowed the analysis of PAHs in samples collected with low volumes (< 0.5 m3), such as punch samples from whole filters or filter strips from personal exposure monitoring equipment. PAHs were successfully analyzed in filter strips from real-time Aethalometer AE51 equivalent black carbon (eBC) analyzers used in urban and rural sites, and in personal exposure monitors of firefighters during prescribed burns. The correlations between PAHs and eBC in these analyses were very strong (r2 ≥ 0.93). However, the equations obtained reflected the dominance of different emission sources, such as traffic in urban areas, wood burning for domestic heating, or wildfires. The method reported here affords the analyses of PAHs in high precision studies of atmospheric PM samples, e.g., high frequency sampling of low volumes, affording personal exposure monitoring assessments.

Effects of Bee Pollen Derived from Acer mono Maxim. or Phellodendron amurense Rupr. on the Lipid Composition of Royal Jelly Secreted by Honeybees.

Royal jelly is a specific product secreted by honeybees, and has been sought after to maintain health because of its valuable bioactive substances, e.g., lipids and vitamins. The lipids in royal jelly come from the bee pollen consumed by honeybees, and different plant source of bee pollen affects the lipid composition of royal jelly. However, the effect of bee pollen consumption on the lipid composition of royal jelly remains unclear. Herein, we examined the influence of two factors on the lipid composition of royal jelly: first, two plant sources of bee pollen, i.e., Acer mono Maxim. (BP-Am) and Phellodendron amurense Rupr. (BP-Pa); secondly, different feeding times. Lipidomic analyses were conducted on the royal jelly produced by honeybees fed BP-Am or BP-Pa using ultra-high performance liquid chromatography (UPLC)-Q-Exactive Orbitrap mass spectrometry. The results showed that the phospholipid and fatty acid contents differed in royal jelly produced by honeybees fed BP-Am compared to those fed BP-Pa. There were also differences between timepoints, with many lipid compounds decreasing in abundance soon after single-pollen feeding began, slowly increasing over time, then decreasing again after 30 days of single-pollen feeding. The single bee pollen diet destroyed the nutritional balance of bee colonies and affected the development of hypopharyngeal and maxillary glands, resulting in differences in royal jelly quality. This study provides guidance for optimal selection of honeybee feed for the production of high-quality royal jelly.

A UPLC Q-Exactive Orbitrap Mass Spectrometry-Based Metabolomic Study of Serum and Tumor Tissue in Patients with Papillary Thyroid Cancer

To find the metabolomic characteristics of tumor or para-tumor tissues, and the differences in serums from papillary thyroid cancer (PTC) patients with or without lymph node metastasis. We collected serums of PTC patients with/without lymph node metastasis (SN1/SN0), tumor and adjacent tumor tissues of PTC patients with lymph node metastasis (TN1 and PN1), and without lymph node metastasis (TN0 and PN0). Metabolite detection was performed by ultra-high performance liquid chromatography combined with Q-Exactive orbitrap mass spectrometry (UPLC Q-Exactive). There were 31, 15, differential metabolites in the comparisons of TN1 and PN1, TN0 and PN0, respectively. Seven uniquely increased metabolites and fourteen uniquely decreased metabolites appeared in the lymph node metastasis (TN1 and PN1) group. Meanwhile, the results indicated that four pathways were co-owned pathways in two comparisons (TN1 and PN1, TN0 and PN0), and four unique pathways presented in the lymph node metastasis (TN1 and PN1) group. Common or differential metabolites and metabolic pathways were detected in the lymph node metastasis and non-metastatic group, which might provide novel ways for the diagnosis and treatment of PTC.

The fate and behavior of glufosinate-enantiomers and their metabolites in open-field soil and weeds

In this study, a chiral method based on high performance liquid chromatography–Q-Exactive Orbitrap Mass Spectrometry was developed to determine glufosinate stereoisomers and three metabolites in weed. Fortified recoveries in weed and soil samples were from 78.6 to 94.3 %, with relative standard deviations of less than 9.8 % and fortified values ranging from 0.04 to 40 mg/kg for the glufosinate enantiomers and 0.08–8 mg/kg for three metabolites. When glufosinate was given at the peak of weed growth in three orchards, it was mostly distributed and degraded in the weeds, with little remaining in the soil. The two glufosinate enantiomers degraded rapidly in the weeds and soils, with half-lives ranging from 0.7 to 3.1 days. The degradation of glufosinate enantiomers in Guizhou and Hunan weeds was enantioselective, with l -glufosinate being preferentially degraded. In Hainan weed, the degradation rate of the two enantiomers was nearly the same. In open field soils, glufosinate enantiomers were almost non-enantioselective. 3-methylphosphinico-propionic acid (MPP) was the primary glufosinate metabolite in weeds and soils, accounting for up to 14 % of the parent. N -acetyl-glufosinate (NAG) was relatively low, with less than 1 % of the parent glufosinate metabolized into 2-methylphosphinico-acetic acid (MPA).

Qualitative Analysis of Multiple Phytochemical Compounds in Tojapride Based on UHPLC Q-Exactive Orbitrap Mass Spectrometry.

Tojapride is composed of Caulis Perillae, Rhizoma Cyperi, Radix Glycyrrhizae, Citrus aurantium L., Coptis chinensis Franch, Pericarpium Citri Reticulatae, Reynoutria japonica Houtt, Tetradium ruticarpum, and Cleistocactus sepium. It has the effects of inhibiting gastric acid and relieving pain. It is clinically used for treating gastroesophageal reflux disease. To further study the pharmacodynamic properties of Tojapride, the systematic characterization of the chemical constituents in Tojapride was investigated using ultra-performance liquid chromatography with Q-Exactive Orbitrap mass spectrometry combined with parallel reaction monitoring for the first time. Eventually, a total of 222 compounds, including flavonoids, alkaloids, and glycyrrhizic acid derivatives, were identified based on the chromatographic retention times, MS/MS2 information, and bibliography data; a total of 218 of these were reported for the first time as being present in Tojapride. This newly developed approach provides a powerful tool for extending our understanding of chemical constituents of Tojapride, which can be further extended to other TCMP composition research.

Identification and comparison of palmitoleic acid (C16:1 n-7)-derived lipids in marine fish by-products by UHPLC-Q-exactive orbitrap mass spectrometry

Palmitoleic acid (POA) is a monounsaturated n-7 fatty acid (16:1 n-7) that is active in modulating cardiovascular and metabolic processes. POA-structured phospholipids, diglycerides, and triglycerides (POA-PL, -DG, -TG) are abundantly present in marine life. In this study, POA and POA-lipids contained in crude lipids in visceral by-products of six seafood products (n = 6 for each sample) were analyzed using ultra-high-performance liquid chromatography Q-exactive orbitrap mass spectrometry (UHPLC-QE/MS). The results showed that sardine contains the highest percentage of POA (12.47 %), followed by bigeye tuna (7.31 %), whereas silver pomfret contains the lowest percentage (5.76 %) in total lipids. Nine POA-PLs, eight POA-DGs, and 64 POA-TGs were detected, the PCA analysis showed that although the molecular composition six fish species was similar, their relative content varied significantly across the species. The correlation heatmap showed critical POA-PL and POA-DG/TG correlation coefficients and revealed several highly correlated ions. The presence and ratio of POA-PL and POA-DG/TG molecular species led to the identification of each fish species. These findings improve the understanding of the composition of POA-lipids contained in marine by-products and provide a reference for their comprehensive exploitation.

Differentiation of three commercial tuna species through Q-Exactive Orbitrap mass spectrometry based lipidomics and chemometrics

Mislabeling and adulteration of tuna are common due to the diminishing of morphological characteristics during processing. The tuna authenticity is now being focused in the seafood supply chain. In this study, the lipid profiles of 3 commercial tuna species (skipjack tuna, bigeye tuna and yellowfin tuna) were investigated via ultra-high performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry (UPLC-Q-Exactive Orbitrap MS). A total of 439 lipid species were identified and semi-quantitated by MS-DIAL. Further biomarkers discovery was carried out by chemometrics, leading to 27 lipids being identified as potential lipid biomarkers. Comparisons to reference standards revealed that lipid biomarkers were effective for discrimination of different tuna species. Interestingly, the proposed lipid biomarkers were all glycerophospholipids, implying that they might be the focus of future study.