Using Ultra-performance Liquid Chromatography Quadrupole Time-of-flight Mass Spectrometry Research Articles

Effect of chemical ingredients B and C in Kansui stir-baked with vinegar on fecal metabolomics in rats

To reveal the toxic mechanism of Kansui stir-baked with vinegar(VEK), conducta comparative study on the metabolites of fecal samples of rats before and after being treated with chemical constituents group B and C(VEKB/VEKC) extracted from VEK by metabolomics approach. The fecal samples of each group were analyzed using ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry(UFLC-Q-TOF-MS). Then the data was processed by principal component analysis(PCA) and partial least square discriminant analysis(OPLS-DA) to screen and identify biomarkers relating to the toxicity of VEK. Besides, t-test was adopted for univariate statistical analysis, so as to study the changes of these biomarkers in drug groups before and after being treated with VEKB/VEKC and explore the effect of VEKB/VEKC on the metabolism of rat feces. Furthermore, the toxic mechanism of VEKB/VEKC was explored based on the results of the metabolic pathway analysis. The results displayed that compared with control group, the metabolism of fecal samples of VEKB and VEKC treated groups show obvious changes, and the VEKB treated group show more significant changes. A total of 16 potential biomarkers and 5 metabolic pathways relating to the toxicity of VEK were found and identified. And the toxicity of VEK might be associated with the disorder of such metabolic pathways as tryptophan metabolism, primary bile acid biosynthesis, amino sugar and nucleotide sugar metabolism, purine metabolism, and degradation of valine, leucine and isoleucine. This study provides a scientific basis for the clinical safety application of VEK.

UPLC fingerprint combined with quantitative analysis of multi-components by single marker for quality assessment of Danshen Injection

Testing and analysis of chemical markers is currently the prevailing approach for quality assessment of traditional Chinese medicines. However,several important issues remain to be addressed,including the trade-off between accuracy and coverage. In this study,in order to give full play to the advantages of their respective methods and provide technical support for more comprehensively and rapidly evaluate the quality of Danshen Injection products,a fingerprint method was coupled with quantitative analysis of multicomponents by single marker( QAMS),with Danshen Injection as the carrier. Ultra performance liquid chromatography( UPLC) was used to establish the quantitative fingerprint. The UPLC fingerprints contained 13 common peaks,11 of which were identified by using ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry( UPLC-QTOF-MS). Furthermore,with sodium danshensu as the internal reference substance,relative correction factors( RCFs) of protocatechuic aldehyde,caffeic acid,rosmarinic acid,lithospermic acid,and salvianolic acid B were calculated through slope analysis method,and the QAMS method was adopted to determine the contents of these 6 components. The UPLC fingerprint was employed to assess the consistency of 12 batches of Danshen Injection,which showed good batch-to-batch consistency with similarity higher than 0. 99. In the comparison of contents of the six constituents obtained by QAMS and external standard method( ESM),RSD was all less than 4. 3%,indicating the good accuracy of the QAMS method. The QAMS method developed in this study combined with UPLC fingerprint can comprehensively reflect the internal quality of Danshen Injection when only the reference substance sodium danshensu is consumed,with greatly reduced detection cost and time. It provides a technical basis for further improving the quality standard of Danshen Injection.

Study on Metabolic Trajectory of Liver Aging and the Effect of Fufang Zhenzhu Tiaozhi on Aging Mice.

The aim of this study was to investigate the metabolic trajectory of liver aging, the effect of FTZ against liver aging in aging mice, and its mechanism using ultraperformance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS). Methods: A total of 80 C57BL/6J Narl mice were randomly divided into five groups: 3-month-old group, 9-month-old group, 14-month-old group, 20-month-old group, and FTZ treatment group (20 months old). The mice in the treatment group received a therapeutic dose of oral FTZ extract (1.0 g/kg, on raw material weight basis) once daily during the experiment. The other groups received the corresponding volume of oral normal saline solution. Liver samples of all five groups were collected after 12 weeks, and UPLC-Q-TOF/MS was used to analyze metabolic changes. Orthogonal partial least squares–discriminant analysis (OPLS-DA) was used to analyze the resulting data. Additionally, cholesterol (TC), triglyceride (TG), aspartate aminotransferase (AST), alanine aminotransferase (ALT), secretion levels of TNF-α, IL-6, 5-LOX, and COX-2, as well as their relative mRNA expression in the liver were determined. Results: The levels of TC, TG, AST, and ALT were increased, and liver tissue structure was damaged. The secretion levels of TNF-α, IL-6, 5-LOX, and COX-2, as well as their relative mRNA expression in the liver also increased with aging. FTZ administration reduced the symptoms of liver aging. The OPLS-DA score plot illustrated the effect of FTZ against liver aging, with N-acetyl-leukotriene E4, 20-hydroxy-leukotriene E4, leukotriene E4, and arachidonic acid among the key biomarkers. The pivotal pathways revealed by pathway analysis included arachidonic acid metabolism and biosynthesis of unsaturated fatty acids. The mechanism by which FTZ reduces the symptoms of liver aging in mice might be related to disorders of the abovementioned pathways. Conclusion: A metabolomic approach based on UPLC-Q-TOF/MS and multivariate statistical analysis was successfully applied to investigate the metabolic trajectory of liver aging. FTZ has a protective effect against liver aging, which may be mediated via interference with the metabolism of arachidonic acid, biosynthesis of unsaturated fatty acids, and downregulation of pro-inflammatory factors in the liver in mice in vivo.

Comparative Analysis of Panax ginseng Berries from Seven Cultivars Using UPLC-QTOF/MS and NMR-Based Metabolic Profiling.

The commercial use of Panax ginseng berries is increasing as P. ginseng berries are known to contain large amounts of ginsenosides, and many pharmacological activities have been reported for the various ginsenosides. For the proper use of P. ginseng berries, it is necessary to study efficient and accurate quality control and the profiling of the overall composition of each cultivar. Ginseng berry samples from seven cultivars (Eumseung, Chung-buk Province, Republic of Korea) were analyzed using ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-QTOF/MS) for profiling of the ginsenosides, and high-resolution magic-angle-spinning nuclear magnetic resonance (HR-MAS NMR) spectroscopy for profiling of the primary metabolites. Comparing twenty-six ginsenoside profiles between the variant representatives and between the violet-stem variant, Kumpoong and Sunwon were classified. In the case of primary metabolites, the cultivars Kumpoong and Gopoong were classified. As a result of correlation analyses of the primary and secondary metabolites, in the Gopoong cultivar, the metabolism was found to lean toward energy metabolism rather than ginsenoside synthesis, and accumulation of osmolytes was low. The Gopoong cultivar had higher levels of most of the amino acids, such as arginine, phenylalanine, isoleucine, threonine, and valine, and it contained the highest level of choline and the lowest level of myo-inositol. Except for these, there were no significant differences of primary metabolites. In the Kumpoong cultivar, the protopanaxatriol (PPT)-type ginsenosides, ginsenoside Re and ginsenoside Rg2, were much lower than in the other cultivars, while the other PPT-type ginsenosides were inversely found in much higher amounts than in other cultivars. The Sunwon cultivar showed that variations of PPT-type ginsenosides were significantly different between samples. However, the median values of PPT-type ginsenosides of Sunwon showed similar levels to those of Kumpoong. The difference in primary metabolites used for metabolism for survival was found to be small in our results. Our data demonstrated the characteristics of each cultivar using profiling data of the primary and secondary metabolites, especially for Gopoong, Kumpoong, and Sunwon. These profiling data provided important information for further research and commercial use.

Development of UPLC-Q-TOF-MS Coupled with Cation-exchange Solid-phase Extraction Method for the Determination of Ten Pyrrolizidine Alkaloids in Herbal Medicines.

Pyrrolizidine alkaloids are secondary metabolites of plants and can cause significant hepatotoxicity in humans. In this study, a fast and simple method was developed to determine ten pyrrolizidine alkaloids (PAs) in six types of herbal medicines using ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS). An efficient solid-phase extraction procedure was carried out by using strong cation-exchange cartridges and the parameters were optimized. The established analytical method was validated and the results showed that the method presented satisfactory accuracy and precision. The established method was successfully applied for the determination of PAs in six herbal plants, including Senecionis Scandentis Hebra, Arnebiae Radix, Asteris Radix Et Rhizoma, Farfarae Flos, Senecionis Cannabifolii Herba and Emilia sonchifolia. PAs were found in all of these herbal plant samples. Eight types of related commercial herbal drugs were also detected, six of them were detected with different amounts of PAs. This work not only provided a powerful technical platform for both qualitative and quantitative analysis of PAs in herbal medicines, but also obtained information concerning PAs in these herbal samples, which could provide reference to the government regulatory authorities and non-governmental organizations.

An integrative investigation on the efficacy of Plantaginis semen based on UPLC-QTOF-MS metabolomics approach in hyperlipidemic mice

Plantaginis semen, the dried mature seed of Plantago asiatica L. or Plantago deprdssa Willd., has a prominent effect on the treatment of obesity, type 2 diabetes and lipid disorders, however, its clinical application is limited due to inadequate in-depth mechanism exploration and incomplete discussion of action targets of its in vivo. Therefore, an untargeted metabolomics approach was firstly applied to study the serum metabolic differences in mice. Metabolomics analysis was performed using ultra performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF-MS) together with multivariate statistical data analysis. The results showed that Plantaginis semen can mainly improve blood lipids, some degree in blood glucose and insulin levels in high-fat mice, in addition, the phenotype of liver and fat stained sections demonstrated remarkable results. A total of 22 metabolites involved in arachidonic acid, glycerophospholipid, glycosphingolipid, linoleate, Omega-3 fatty acid, phosphatidylinositol phosphate and tyrosine metabolisms were identified. In further, it was found that the possible mechanisms of Plantaginis semen on hyperlipidemic mice lied in the biosynthesis of thyroxine, biological effects of enzymes of phospholipase A2 activity, glucosylceramide synthase and inositol essential enzyme 1α, genes expressions of fatty acid metabolism and inflammation. Serum metabolomics revealed that Plantaginis semen could cure the organism disease via regulating multiple metabolic pathways which will be helpful for understanding the mechanism of this herb and providing references for better applications of it in clinic, even researches on other TCMs.

Anti-inflammatory and antioxidative effects of Dan-Lou tablets in the treatment of coronary heart disease revealed by metabolomics integrated with molecular mechanism studies

Ethnopharmacological evidenceThe Dan-Lou tablet (DLT), a well-known Chinese prescription, has definitive clinical efficacy in the treatment of precordial discomfort and pain caused by coronary heart disease (CHD). However, the pharmacological mechanism of DLT in the treatment of CHD has not been clearly elucidated and needs to be further explored. Aim of the studyWe aimed to identify relevant biological pathways by assessing changes in biomarkers in response to DLT intervention in CHD to reveal the potential biological mechanism of DLT treatment for CHD. Materials and methodsThe major chemical components in DLT were qualitatively analyzed using ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS), and a model of CHD in rats was subsequently established with a high-fat diet and left anterior coronary artery ligation (LADCA) followed by DLT intervention. Next, the metabolic profile of rat serum samples was analyzed using nontargeted metabolomics, wherein changes in the metabolites in serum samples before and after DLT administration were measured by PLS-DA, and two pathways of DLT treatment for CHD were predicted. Finally, predicted metabolomic pathways were verified by detecting and analyzing tissues from the rat model, revealing the mechanism of DLT in the treatment of CHD. ResultsForty-five major chemical components were identified by the chemical characterization of DLT. In terms of metabolism, 17 biomarkers of CHD in rats were identified. Among these biomarkers, linoleic acid, γ-linolenic acid and lysophosphatidylcholines (LPCs) were found to play an important role in energy metabolism and glycerophospholipid metabolism. Protein analysis revealed that EGFR phosphorylation was inhibited in CHD rats after DLT treatment, which lowered the expression of TNF-α, IL-6 and MMP9, decreased the expression levels of ox-LDL and MDA, and increased the expression of SOD. ConclusionThe mechanism of DLT in the treatment of CHD involves inhibiting the expression of EGFR and the activation of the MAPK signaling pathway by regulating glycerophospholipid metabolism (LPCs) and energy metabolism (linoleic acid and γ-linolenic acid). Therefore, inflammation-related (TNF-α, IL-6, MMP9) and oxidative stress-related (ox-LDL, MDA, SOD) indicators are affected, leading to the regulation of the oxidative stress state and anti-inflammatory effects.

Determination of multiple chelator complexes in aqueous matrices using ultra-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry

The determination of aminopolycarboxylate chelators in environmental samples has remained an analytical challenge due to the structural similarities of these species and their minute concentrations in such matrices. Herein, we report a fast and sensitive technique for the determination of multiple chelator complexes in an aqueous matrix using ultra-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Eight chelators, including non-biodegradable (EDTA, EDTAOH, GEDTA, DPTAOH and DTPA) and biodegradable (EDDS, GLDA, and MGDA) variants were examined after complexation with CuII. The detection of these species using reverse-phase chromatography was compared with that achieved with hydrophilic interaction chromatography based on the corresponding peak resolution and retention time. The effect of varying the composition and pH of the mobile phase on the corresponding peak profiles and intensities for the chelator complexes was also evaluated. The CuII-derivatives of the chelators were individually detected under the optimized operating conditions. Relative to high-performance liquid chromatography equipped with a photodiode array detector, the developed UPLC-Q-TOF-MS technique provides rapid determination of chelator complexes in aqueous matrices with high sensitivity and superior peak resolution. The limit of detection ranged from 1.7–36 nmol L–1, and the limit of quantification ranged from 5.7–120 nmol L–1 for the eight chelator complexes in solution. The coefficients of determination (R2) were 0.962–0.999 for the chelators with an average relative uncertainty of 2.2%. The method was validated using a simulated mixed matrix and river water by standard addition (recovery: 83−100%).

Clinical metabolomics analysis of therapeutic mechanism of Tongmai Yangxin Pill on stable angina

Tongmai Yangxin Pill (TMYX) is a traditional Chinese medicine for the treatment of angina and arrhythmia. Although its clinical application is extensive, and the curative effect is significant, little information is available on the molecular biological basis and therapeutic mechanism of TMYX for the treatment of stable angina. In this study, we analyzed serum samples of clinical patients collected from seven different clinical units in China after oral administration of TMYX using ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS). Multiple statistical analysis including principal component analysis (PCA) and partial least square discrimination analysis (PLS-DA), were used to examine metabolite profile changes in serum samples. After TMYX treatment, 10 biomarkers were reversed to the normal conditions. The above biomarkers were mainly involved in energy metabolism, amino acid metabolism, oxidative stress and inflammation. These results suggested that TMYX exerted therapeutic effects by improving myocardial energy supply disorder and amino acid dysfunction, and attenuating oxidative stress and inflammation. The present study, as the first multicenter clinical study which reveals the molecular biological basis and therapeutic mechanism of TMYX on stable angina, can provide objective indicators for efficacy evaluation of TMYX on stable angina. And it also lays a foundation for the use of TMYX clinically.

Detection of the designer benzodiazepine flunitrazolam in urine and preliminary data on its metabolism.

Designer benzodiazepines have emerged as recreational drugs. They are available via the Internet without control and are found in the form of falsified (fake) medicines. For some of them, limited information concerning their effects, their toxicity, and their detection in bio fluids is available in the literature. For others, nothing has been published, as in the case of flunitrazolam (FNTZ). To gain preliminary data on its elimination parameters in urine and to investigate its metabolism, one of the authors ingested one pink tablet bought on the Internet, after confirming the absence of other compounds and agreement with the labeled dosage (0.25mg) by nuclear magnetic resonance (NMR). A software algorithm (MetaboLynx, Waters, Milford, MA, USA) was used to predict FNTZ biotransformation and four potential metabolites were proposed: 4-hydroxy-FNTZ, desnitro-FNTZ, 7-amino-FNTZ, and 7-acetamido-FNTZ. Urine samples were collected over 72hours following oral administration of one tablet. After liquid/liquid extraction at pH9.5, FNTZ concentrations were determined using ultra performance liquid chromatography-triple quadrupole-mass spectrometry (UPLC-QqQ-MS/MS). FNTZ remained detectable in hydrolyzed urine for 21hours after ingestion, with concentrations ranging between 1 and 18ng/mL. About 3% of the initial dose was excreted in urine as total unchanged FNTZ during this period. In vitro experiments (HLM incubations) were performed using ultra performance liquid chromatography-quadrupole time of flight-mass spectrometry (UPLC-QTOF-MS) in order to investigate the potential CYP- and UGT-dependent metabolites where only 7-amino-FNTZ was detected as the only metabolite. However, in the urine specimens, desnitro-FNTZ, 7-acetamido-FNTZ and 7-amino-FNTZ were the main detected compounds. The identification of FNTZ metabolites dramatically improves the detection windows of the drug up to 37hours.

Metabolomic profiling of rat urine after oral administration of the prescription antipyretic Hao Jia Xu Re Qing Granules by UPLC/Q-TOF-MS.

Hao Jia Xu Re Qing Granules (HJ), is an effective clinically used antipyretic based on traditional Chinese medicine. Although its antipyretic therapeutic effectiveness is obvious, its therapeutic mechanism has not been comprehensively explored yet. In this research, we first identified potential biomarkers which may be relevant for the antipyretic effect of HJ based on urine metabolomics using ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). A rat model of fever was established using the yeast-induced febrile response. Total-ion-current metabolic profiles of different groups were acquired and the data were processed by multivariate statistical analysis-partial least-squares discriminant analysis. As envisioned, the results revealed changes of urine metabolites related to the antipyretic effect. Fourteen potential biomarkers were selected from the urine samples based on the results of Student's t-test, "shrinkage t", variable importance in projection and partial least-squares discriminant analysis. N-Acetylleucine, kynurenic acid, indole-3-ethanol, nicotinuric acid, pantothenic acid and tryptophan were the most significant biomarkers found in the urine samples, and may be crucially related to the antipyretic effect of HJ. Consequently, we propose the hypothesis that the significant antipyretic effect the HJ may be related to the inhibition of tryptophan metabolism. This research thus provides strong theoretical support and further direction to explain the antipyretic mechanism of HJ, laying the foundation for future studies.

Untargeted metabolomics approach (UPLC-Q-TOF-MS) explores the biomarkers of serum and urine in overweight/obese young men.

Obesity is linked to metabolic diseases characterized by insulin resistance, such as diabetes and cardiovascular disease. In this study, we investigated the metabolic disorders of uncomplicated obesity to identify early alterations in biological systems. Metabolic differences between overweight/obese (n=36) and normal-weight (n=35) young Chinese men without known metabolic disorders were assessed. Metabolic profiling of the serum and urine was performed using ultra-performance liquidchromatography quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Partial least squares discriminant analysis (PLS-DA) was undertaken to reveal and classify the differences between the two groups. Compared to normal-weight men, obese men had higher levels of the serum metabolites phenylalanine, Phe-Phe, and L-tryptophan, whereas those of p-cresol sulfate and p-cresol were less in obesity. Urinary metabolites phenylacetamide, L-glutamine, phenylacetylglutamine, indoxyl sulfate, p-cresol, and p-cresol sulfate were greater in obese men. These findings indicate that disorders involving aromatic amino acids and the tricarboxylic acid cycle (TCA) have microbiomic involvement in the uncomplicated phase of obesity.

Anti-aging potential of extracts from Sclerocarya birrea (A. Rich.) Hochst and its chemical profiling by UPLC-Q-TOF-MS

BackgroundDegradation of components of the extracellular matrix such as elastin and collagen by elastase and collagenase accelerates skin aging. Phytochemicals that inhibit the activity of these enzymes can be developed as anti-aging ingredients. In this study, an investigation of the anti-aging properties of Sclerocarya birrea (A. Rich.) Hochst (Marula) extracts was conducted in vitro with the aim of developing chemically characterized anti-aging ingredients.MethodsMarula stems, leaves and fruits were extracted using methanol:dichloromethane (DCM) (1:1). The stems were later extracted using acetone, ethanol, methanol:DCM (1:1) and sequentially using hexane, DCM, ethyl acetate and methanol. The stem ethanol extract was defatted and concentrated. Elastase and collagenase inhibition activities of these extracts and Marula oil were determined using spectrophotometric methods. The chemical profile of the ethanolic stem extract was developed using Ultra-performance-liquid chromatography quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS) with MassLynx software. Pure standards were used to confirm the identity of major compounds and were screened for anti-elastase and anti-collagenase activity.ResultsMarula stems extracts were the most active as they exhibited anti-elastase activity comparable to that of elafin (> 88%) and anti-collagenase activity as potent as EDTA (> 76%). The leaf extract had moderate anti-elastase activity (54%) but was inactive agains collagenase. Marula fruits and oil exhibited limited activity in both assays. The ethanolic extract of Marula stems was the most suitable based on its acceptability to the cosmetic industry and its anti-collagenase activity (99%). Defatting and concentration improved its antiaging activity and lowered the colour intensity. Six compounds have been tentatively identified in the chemical profile of the ethanolic extract of Marula stems of which four; quinic acid, catechin, epigallocatechin gallate and epicatechin gallate have been confirmed using pure standards. Epigallocatechin gallate and epicatechin gallate were as potent (p < 0.05) as EDTA at 5 μg/ml in the anti-collagenase assay.ConclusionsThe ethanolic extract of Marula stems can be developed into an anti-aging ingredient as it exhibited very good in vitro anti-aging activity and its chemical profile has been developed. Epicatechin gallate and epigallocatechin gallate contribute to the anti-aging activity of Marula stem ethanol extract.

Metabolic analyses reveal different mechanisms of leaf color change in two purple-leaf tea plant (Camellia sinensis L.) cultivars

Purple-leaf tea plants, as anthocyanin-rich cultivars, are valuable materials for manufacturing teas with unique colors or flavors. In this study, a new purple-leaf cultivar “Zixin” (“ZX”) was examined, and its biochemical variation and mechanism of leaf color change were elucidated. The metabolomes of leaves of “ZX” at completely purple, intermediately purple, and completely green stages were analyzed using ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF-MS). Metabolites in the flavonoid biosynthetic pathway remained at high levels in purple leaves, whereas intermediates of porphyrin and chlorophyll metabolism and carotenoid biosynthesis exhibited high levels in green leaves. In addition, fatty acid metabolism was more active in purple leaves, and steroids maintained higher levels in green leaves. Saponin, alcohol, organic acid, and terpenoid-related metabolites also changed significantly during the leaf color change process. Furthermore, the substance changes between “ZX” and “Zijuan” (a thoroughly studied purple-leaf cultivar) were also compared. The leaf color change in “Zijuan” was mainly caused by a decrease in flavonoids/anthocyanins. However, a decrease in flavonoids/anthocyanins, an enhancement of porphyrin, chlorophyll metabolism, carotenoid biosynthesis, and steroids, and a decrease in fatty acids synergistically caused the leaf color change in “ZX”. These findings will facilitate comprehensive research on the regulatory mechanisms of leaf color change in purple-leaf tea cultivars.

Identification of metabolites of Chenxiang Huaqi pill in rats based on UPLC-Q-TOF-MS

To analyze the metabolites of Chenxiang Huaqi pill in rats by using ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS). The separation was performed on Phenomenex Kinetex C₁₈ column, with the acetonitrile -0.1% formic acid as the mobile phase for gradient elution at a flow rate of 0.8 mL·min⁻¹. The data were collected by the positive ion mode of ESI source. The plasma and urine total ion chromatograms of the rats in blank group and treatment group were used to analyze the targeted ion chromatograms. The results showed that 24 compounds were detected in the plasma and urine, including 5 prototype components and 19 metabolites. The major metabolic pathways included hydration, glucuronidation, demethylation, hydrolysis, hydroxylation and sulfation. The method was rapid, simple and sensitive, and can be used to rapidly identify the metabolites of Chenxiang Huaqi pill that can be absorbed in rats, providing a reference for the study of the absorption and metabolism mechanism of Chenxiang Huaqi pill in vitro.

Urine and plasma metabolomics study on potential hepatoxic biomarkers identification in rats induced by Gynura segetum

Ethnopharmacological relevanceGynura segetum (GS) is an herbal medicine containing Pyrrolizidine Alkaloids (PAs) that causes hepatic sinusoidal obstruction syndrome (HSOS). Aim of the studyTo discover potential biomarkers and metabolic mechanisms involved in the hepatotoxicity induced by GS. MethodsSD rats were randomly divided into 4 groups including Saline, the decoction of GS high, medium and low dosage at dosages of 3.75g • kg−1, 7.5g • kg−1 and 15g • kg−1. A metabolomics approach using Ultraperformance Liquid Chromatography -Quadrupole-Time-of-Flight / Mass Spectrometry (UPLC-Q-TOF/MS) was developed to perform the plasma and urinary metabolic profiling analysis, and identified differential metabolites by comparing the saline control group and decoction of GS groups. ResultsThe herbal was presented dosage-dependent led to ingravescence of hepatotoxicity after the rats were consecutively given with the decoction of GS at varied dosages. A total of 18 differential metabolites of decoction of GS-induced hepatotoxicity were identified, while 10 of them including arginine, proline, glutamate, creatine, valine, linoleic acid, arachidonic acid, sphinganine, phytosphingosine, and citric acid could be discovered in urine and plasma, and primarily involved in Amino acid metabolism, Lipids metabolism and Energy metabolism. ConclusionsThe results suggested that the differential metabolites of arginine, creatine, valine, glutamine and citric acid were verified as potential markers of GS-induced hepatotoxicity via the regulation of multiple metabolic pathways primarily involving in Amino acids metabolism and Energy metabolism.

Postoperative Delirium Development Associated with Metabolic Alterations Following Hemi-Arthroplasty in Elderly

Background: Postoperative delirium (POD) is a common and harmful complication following surgery in elderly and possible link between metabolic changes during perioperative period and POD development is yet to be investigated. Methods: A nested case-control study was performed through metabolomics to find out the metabolic abnormalities between patients with and without POD. Elderly patients with hip fracture who had hemi-arthroplasty were recruited and their delirious states were assessed for three days after surgery. Simultaneously, fasting blood samples were collected on the morning of surgery and on the first postoperative day. Ultimately, 244 elderly patients who met the inclusion and exclusion criteria completed our observations. The blood samples from 60 patients with POD and 60 matched controls were analyzed using ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) and gas chromatography time-of-flight mass spectrometry (GC-TOF-MS). Findings: Sixty patients (24.6%) developed POD. Eighteen metabolites differed between the 2 groups before surgery, and twenty-three metabolites differed between the 2 groups after surgery. Similarly, thirty metabolites in POD group differed before and after surgery, and thirty-two metabolites in NPOD (Non-POD) group differed before and after surgery. After retrieved KEGG database, our data indicated that, in POD group, ω3 and ω6 fatty acids were lower than NPOD group at both before and after surgery, intermediates of tricarboxylic cycle were lower than NPOD group but products of glycolysis were higher than NPOD group after surgery, branched-chain amino acids (BCAAs) / aromatic amino acids (AAAs) ratio was lower than NPOD group after surgery. Interpretation: The metabolic abnormalities, including deficiencies of ω3 and ω6 fatty acids, abnormal oxygen metabolism, metabolic imbalances in aromatic and branched-chain amino acids, may potentially diminish the brain's inherent protective capacity and hence could contribute to the POD development. Funding Statement: This work was supported by grants from the Shanghai Municipal Commission of Health and Family Planning Foundation for Key Developing Disciplines (2015ZB0103) and the Shanghai Jiao Tong University Biomedical Engineering Cross Research Foundation (YG2015MS15). Declaration of Interests: The authors report no conflicts of interest. Ethics Approval Statement: The study was approved by the Ethics Committee of Shanghai Sixth People’s Hospital affiliated with Shanghai Jiao Tong University (Shanghai, China) and was registered (ChiCTR-CPC-15006141). Written informed consent was obtained from all the participants prior to the study.

Plasma metabolic profiling analysis of Cortex Periplocae-induced cardiotoxicity based on UPLC/Q-TOF-MS.

Cortex Periplocae is a well-known form of traditional medicine with its unique cardiotonic action, anti-tumor activity and immune regulation effect. However, improper use of Cortex Periplocae often leads to cardiac toxicity, which in the most severe cases can even be life-threatening. Biochemical tests and histopathological examinations are primary methods for clinical trials. However, such approaches are time-consuming, lack specificity and have low sensitivity, which can easily lead to negative results in studies. Therefore, a more scientific and systematic evaluation of Cortex Periplocae cardiotoxicity is particularly important. In this study, we established a method that combines metabonomics with trend analysis of a gavage concentration series to find cardiac toxicity biomarkers of Cortex Periplocae. We created rat cardiotoxicity models, in which the toxicity was caused by Cortex Periplocae. We collected data from rat plasma samples based on metabonomics using ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS). Multiple statistical analyses, such as principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA), were used to examine metabolite profile changes in plasma samples to screen potential cardiotoxicity biomarkers and metabolic pathways. Compared with the control group, after 7 days administration, the pathological sections showed cardiac toxicity. Moreover, some metabolites in the body changed significantly. Receiver operating characteristic curve (ROC) analysis showed that there are 11 metabolites related with cardiac toxicity, which play a role in “phenylalanine, tyrosine and tryptophan biosynthesis”; “phenylalanine metabolism”; “valine, leucine and isoleucine biosynthesis”; “glycerophospholipid metabolism” as well as “pantothenate and CoA biosynthesis”. These metabolites can better explain the cardiotoxicity mechanism of Cortex Periplocae and provide a scientific and systematic method to evaluate the cardiotoxicity of Cortex Periplocae.

Rapidly identify chemical constituents of Schisandra propinqua by UPLC-Q-Tof/MSE

This study was to identify the chemical constituents of Schisandra propinqua, one herbal medicine of Yi nationality in China by using ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF/MSE). Acetonitrile- water containing 0.1% formic acid was used as the mobile phase for gradient elution. Data were collected under ESI negative mode and ESI positive mode, and then screened and verified by the software of UNIFI and Masslynx4.1. Based on the accurate mass, fragment ions, neutral losses, mass error, retention time, reference substance, isotope information, the intensity of fragments, as well as the previous reports, the known compounds were validated and identified. The chemical structures of the unknown components were identified according to exact molecular weight, MS fragment, chromatographic retention behavior, and characteristic fragments of known congener compounds. A total of 68 chemical components were identified from S. propinqua, including 3 flavonoids, 10 flavanols, 34 lignans compounds (including 20 dibenzocyclooctene lignans), 4 triterpenoids, 17 organic acid and other compounds. 37 compounds of them were found in S. propinqua for the first time, and one potential compound needed to be identified.

Occurrence, Distribution and Health Risk of the Phthalate Esters in Riparian Soil in the Fluctuating Zone of the Three Gorges Reservoir

Soil samples were collected in the fluctuating zone of the Three Gorges Reservoir to analyze the concentrations and distribution of phthalate esters (PAEs) using ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS). Meanwhile, the interrelationships among the PAE congeners were analyzed and the potential sources of PAEs in soil were determined by the correlation analysis and principal component analysis. The results showed that the concentrations of ∑PAEs ranged from 322.0-737.3 ng·g-1 dry weight, with a mean value of 497.2 ng·g-1 dry weight, which lies at the low end of the worldwide Figures. DIBP was the most abundant PAEs congener, followed by DBP and DEHP, contributing to 94.5% of the total PAEs. However, DEHP was not the main pollutant in this study compared to other studies, which might be ascribed to the release of DEHP from the flooded soil to the surrounding water but the degradation by microorganisms in the soil cannot be completely ruled out. Significant interrelationships were observed among DMP, DEP, DBP, and DIBP. The principal component analysis showed that DMP, DEP, DBP, and DIBP may have similar sources. In this study, the daily intake dose was lower than the reference dose (RfD) proposed by the United States Environmental Protection Agency (USEPA), and the carcinogenic risk of six PAE congeners was far below the tolerable values proposed by the USEPA. Nevertheless, the daily intake dose of DBP accounted for 60% to 84% of total PAEs; thus, more attention should be paid to the potential health risks of DBP.