Ultra-performance Liquid Chromatography Quadrupole Research Articles

Illuminating the hepatotoxic mechanism of norcantharidin in rats using metabolomics analysis

BackgroundNorcantharidin (NCTD) has multiple antitumor effects. However, NCTD can induce significant hepatotoxicity and the mechanism of hepatotoxicity is not clear for now.ObjectiveThis study aimed to explore the hepatotoxicity of NCTD in rat by ultra-performance liquid chromatography (UPLC) quadrupole time-of-flight (Q-TOF)-MS (UPLC/Q-TOF-MS) metabolomics.ResultsSerum biochemical indices including alanine aminotransferase (ALT) and total bilirubin (T-BIL) were significantly increased. Histopathological and ultrastructure results revealed that hepatocytes were damaged. Furthermore, the metabolomics results showed that 11 metabolites in serum and 8 metabolites in liver were differential metabolites for NCTD hepatotoxicity. Four metabolic pathways including the sphingolipid metabolism, purine metabolism, arachidonic acid metabolism, and glycerophospholipid metabolism were the key metabolic pathways related to NCTD hepatotoxicity.ConclusionThe metabolomics analysis in this study reveal new clues on the hepatotoxicity mechanism of NCTD in rats. These findings have potential applications in the toxicity study of NCTD.

Bioassay and UPLC-Q-Orbitrap-MS/MS guided isolation of polycyclic polyprenylated acylphloroglucinols from St. John's wort and their neuroprotective activity

St. John’s wort ( Hypericum perforatum L.) is a popular dietary supplement ingredient used for the treatment of mild-to-moderate depression in the United States and Germany. Reported studies mainly focused on the biological evaluation and mechanism study of its crude extracts or two main components (namely, hypericin and hyperforin). However, it is unclear whether other phytochemicals including polycyclic polyprenylated acylphloroglucinols (PPAPs) contributed to the neuroprotective effects of H. perforatum. Here, bioassay and ultra performance liquid chromatography–quadrupole orbitrap mass spectrometry (UPLC-Q-Orbitrap-MS/MS) guided isolation were applied to discover bioactive PPAPs from the crude extracts of H. perforatum. A new PPAP, named hyperforen A ( 2 ), along with nine known PPAPs ( 1 and 3 – 10 ), were identified from an ethyl acetate extract of H. perforatum . To the best of our knowledge, compound 2 represents the first PPAP with an unprecedented bicyclo[7.3.0]dodecane core. The chemical structures of the isolates were elucidated by extensive spectroscopic analysis and electronic circular dichroism calculations. Moreover, compounds 1 – 3 , 6 , and 10 (10 μM) exhibited significant neuroprotective effects against corticosterone-induced injury in PC12 cells. Fingings from the current study suggest that bioactive PPAPs from H. perforatum are promising compouds for the management of depression.

Rapid identification of chemical components in Zhizi Baipi decoction by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry coupled with a novel informatics UNIFI platform.

Zhizi Baipi decoction is a classic traditional Chinese medicine formula for the treatment of jaundice and various liver diseases. The chemical components of Zhizi baipi decoction were not clear resulting of the paucity of relevant studies, which hindered the elucidation of the pharmacological mechanism, and the comprehensive development and utilization of Zhizi baipi decoction in clinical. In this study, ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry combined with the UNIFI natural product information analysis platform was used to rapidly analyze and identify the chemical components in Zhizi baipi decoction. A total of 122 chemical components, including 53 flavonoids, 16 alkaloids, 25 terpenoids, five phenylpropanoids, 14 organic acids, and seven others, were identified from Zhizi baipi decoction. These compounds may be the active components of Zhizi baipi decoction. The method established in this study can systematically, rapidly, and accurately resolve the chemical components in Zhizi baipi decoction, which lays the foundation for further establishment of the pharmacodynamic substance basis and quality control of Zhizi baipi decoction.

Rapid characterization of chemical constituents in Naoling Pian by LC-MS combined with data processing techniques.

Naoling Pian is a prescription composed of 15 herbs, which is mainly used for the treatment of insomnia in clinical practice. However, the chemical constituents in Naoling Pian are numerous and unclear, which hinders the interpretation of its bioactive constituents and the subsequent research on the material basis for pharmacodynamics. The purpose of this study is to develop a rapid method for identifying the chemical constituents of Naoling Pian using high-throughput ultra-performance liquid chromatography quadrupole time of flight coupled with mass spectrometry combined with a software platform for data processing. The whole composition of Naoling Pian was characterized in positive and negative ion modes. In this experiment, an overall total of 201 constituents were identified by using reference standards, online and self-built databases matching, fragmentation rules analysis of mass spectrometry peaks with a software platform. Meanwhile, Naoling Pian was analyzed for the first time using liquid chromatography-mass spectrometry method, the constituents could be identified in a quick and accurate manner, and the results could provide a scientific basis for the follow-up research on the pharmacodynamic material basis and quality control of Naoling Pian.

Network of the transcriptome and metabolomics reveals a novel regulation of drought resistance during germination in wheat.

The North China Plain, the highest winter-wheat-producing region of China, is seriously threatened by drought. Traditional irrigation wastes a significant amount of water during the sowing season. Therefore, it is necessary to study the drought resistance of wheat during germination to maintain agricultural ecological security. From several main cultivars in the North China Plain, we screened the drought-resistant cultivar JM47 and drought-sensitive cultivar AK58 during germination using the polyethylene glycol (PEG) drought simulation method. An integrated analysis of the transcriptome and metabolomics was performed to understand the regulatory networks related to drought resistance in wheat germination and verify key regulatory genes. Transcriptional and metabolic changes were investigated using statistical analyses and gene-metabolite correlation networks. Transcript and metabolite profiles were obtained through high-throughput RNA-sequencing data analysis and ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry, respectively. A total of 8083 and 2911 differentially expressed genes (DEGs) and 173 and 148 differential metabolites were identified in AK58 and JM47, respectively, under drought stress. According to the integrated analysis results, mammalian target of rapamycin (mTOR) signalling was prominently enriched in JM47. A decrease in α-linolenic acid content was consistent with the performance of DEGs involved in jasmonic acid biosynthesis in the two cultivars under drought stress. Abscisic acid (ABA) content decreased more in JM47 than in AK58, and linoleic acid content decreased in AK58 but increased in JM47. α-Tocotrienol was upregulated and strongly correlated with α-linolenic acid metabolism. The DEGs that participated in the mTOR and α-linolenic acid metabolism pathways were considered candidate DEGs related to drought resistance and the key metabolites α-tocotrienol, linoleic acid and l-leucine, which could trigger a comprehensive and systemic effect on drought resistance during germination by activating mTOR-ABA signalling and the interaction of various hormones.

Profiling the role of microorganisms in quality improvement of the aged flue-cured tobacco

BackgroundThe aging process in the tobacco production, as in other food industries, is an important process for improving the quality of raw materials. In the spontaneous aging, the complex components in flue-cured tobacco (FT) improve flavor or reduce harmful compounds through chemical reactions, microbial metabolism, and enzymatic catalysis. Some believed that tobacco-microbe played a significant part in this process. However, little information is available on how microbes mediate chemical composition to improve the quality of FT, which will lay the foundation for the time-consuming spontaneous aging to seek ways to shorten the aging cycle.ResultsComparing aged and unaged FT, volatile and non-volatile differential compounds (DCs) were multi-dimensionally analyzed with the non-targeted metabolomes based on UPLC-QTOP-MS (the ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry), GC–MS (gas chromatography-mass spectrometer) assisted derivatization and HP-SPME-GC/MS (headspace solid-phase micro-extraction assisted GC–MS). Products associated with the degradation pathways of terpenoids or higher fatty acids were one of the most important factors in improving FT quality. With the microbiome, the diversity and functions of microbial flora were analyzed. The high relative abundance function categories were in coincidence with DCs-related metabolic pathways. According to the correlation analysis, Acinetobacter, Sphingomonas and Aspergillus were presumed to be the important contributor, in which Aspergillus was associated with the highest number of degradation products of terpenoids and higher fatty acids. At last, the screened Aspergillus nidulans strain F4 could promote the degradation of terpenoids and higher fatty acids to enhance tobacco flavor by secreting highly active lipoxygenase and peroxidase, which verified the effect of tobacco-microbes on FT quality.ConclusionsBy integrating the microbiome and metabolome, tobacco-microbe can mediate flavor-related substances to improve the quality of FT after aging, which provided a basis for identifying functional microorganisms for reforming the traditional spontaneous aging.

High-resolution quadrupole time-of-flight mass spectrometry, a powerful technique to include in analytical scheme for the identification of drugs and chemicals in seizures: A case study

A seizure containing 37 items was analyzed with laboratory identification scheme for drugs and chemicals in seized products applying accredited methods based on gas chromatography mass spectrometry (GC-MS) and liquid chromatography with photodiode-array detection (UPLC-PDA), a category A technique combined with category B and C techniques. No identification was obtained for 1 item [analogue of lysergic acid diethylamide (LSD)] and none detected or identified by GC-MS and/or UPLC-PDA for 8 items. All items were subsequently analyzed using ultra performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-Tof) to investigate whether it is appropriate to include this technique in laboratory identification scheme for seized materials. The seizure consisted of individual packages containing blotter papers, capsules and powders, respectively. The powders were present in sealed alu-bags labeled ‘not for human consumption’ and Falcon® tubes, presumably purchased as ‘research chemical’. Methanolic solutions were analyzed. The GC-MS method was described elsewere (Coopman V. Toxicol Anal Clin 2018;30:75–79). Automated library search was performed against NIST, SWGDRUG and CAYMAN mass spectral library and mass spectra published in scientific literature were consulted. The UPLC-PDA analysis was performed with a Waters Acquity UPLC H-Class system. The compounds were separeted on an Acquity UPLC C18 column using gradient elution with a mobile phase consisting of 0.1% (v/v) formic acid aqueous solution and 0.1% (v/v) formic acid in acetonitrile, and photodiode-arra y detection. Automated library search was performed with Empower™ 3 software against in-house and Waters library. A Waters Acquity UPLC I-Class with Xevo G2-XS QTof was used, equipped with UNIFI® scientific information system and Forensic Toxicology Screening Application from Waters with data acquisition parameters for accurate mass analysis in positive and negative mode. Analogues of LSD (1P-LSD, ETH-LAD, AL-LAD) and a designer benzodiazepine (clonazolam) respectively, were identified in blotter paper by UPLC-PDA, GC-MS and confirmed by UPLC-Q-Tof. The unknown analogue of LSD was identified as ALD-52 by UPLC-Q-Tof. 4-fluoromethylphenidate (4F-MPH), N-ethyldeschloroketamine (O-PCE) and mirtagynine were identified in powder by UPLC-PDA, GC-MS and UPLC-Q-Tof. O-PCE was not present in the Forensic Toxicology Screening Application library and was therefore not automatically identified. Salicylic acid, caffeine, trazodone, pseudoephedrine, melatonin and ETH-LAD respectively, were identified in tablet or capsule with routine laboratory identification scheme and confirmed by UPLC-Q-Tof. In a capsule found to contain pseudoephedrine and unknown, a library hit was found for the designer drug N-methyl-2-aminoindane (NM-2AI) by UPLC-Q-Tof. Laboratory identification scheme for the identification of drugs and chemicals in seizures is based on GC-MS and UPLC-PDA, a category A technique and category B, C techniques (SWGDRUG recommendations). Not all drugs and chemicals were detected with both techniques. When identification is obtained with one or any of the applied techniques, a more extensive analysis is recommended. UPLC-Q-Tof is a sensitive and powerful technique, appropriate to include in laboratory identification scheme for seized materials when identification is obtained with one or any of the applied techniques or if the presence of low concentration, a NPS or designer drugs is suspected, allowing confirmation or identification.

Lipidomic analysis reveals the effect of passive smoking on facial skin surface lipid in females

BackgroundSmoking has toxic effects on the skin and can damage it. However, few studies have focused on the lipid profile changes of facial skin surface lipids (SSL) by passive smoking. MethodA cross-sectional analytical study was conducted on middle-aged females volunteered from Henan, China to participate in the study. A total of 20 passive smoking females and 20 non-passive smoking females were recruited for this study. The components of skin surface lipids were measured by ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF-MS). Multivariate data analysis and enrichment analysis were used to investigate the differences in facial SSL between passive and non-passive smoking females. ResultThere were 1247 lipid entities identified in facial SSL between passive and non-passive smoking females. Significant differences in composition of facial SSL were observed between the two groups. After multivariate data analysis suggested, 28 significantly different lipids were identified and classified into four classes in SSL of the female cheeks. As well as 32 significantly different lipids were obtained in SSL of the female foreheads, which included three classes of lipids. Subsequent analysis revealed that the content of fatty acids (FA) in passive smoking females was significantly reduced and the content of glycerolipids (GL) and sphingolipids (SP) increased, compared with the control group. ConclusionThese results indicated that an increase in GLs and SPs of facial lipids and a decrease in FAs in passive smoking females. These changes in lipids might be associated with oxidative stress and interference with signaling pathways by substances in smoke. And passive smoking affected facial SSL and changed the content and metabolism of skin lipids.

Rapid screening of hepatotoxic components in Uncariae Ramulus Cum Uncis based on “component-target-pathway” network

As a multi-base source traditional Chinese medicine, the hepatotoxicity of Uncariae Ramulus Cum Uncis (URCU) has been reported repeatedly in recent years. The lack of clarity of toxic components and toxicity mechanisms is a key issue that needs to be addressed. In this article, a “component-target-pathway” network strategy was established to firstly predicting the hepatotoxic components and the toxicity mechanism of URCU. Ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) and data post-processing technology were used to classify and identify the main components in Uncaria rhynchophylla (Miq.) Miq. ex Havil. (UR) and Uncaria sinensis (Oliv.) Havil. (US). Then, the potential hepatotoxic components were screened by network pharmacology and molecular docking. As a result, 40 components and 39 ingredients were identified in UR and US, respectively. Cadambine, rhynchophylline, corynoxeine, isocorynoxeine, strictosamide and mitraphylline were screened as the potential hepatotoxic ingredients contained both in UR and US. The network pharmacology showed that the potential hepatotoxic components could affect the IL-17 signaling pathway by regulating related targets such as MAPK1 and MAPK14, which might lead to the occurrence of liver injury. This study not only provided a reasonable strategy for the rapid screening of hepatotoxic components in URCU, but also supplied reference and guidance for the rational clinical application and scientific supervision of URCU.

Explore the Potential Ingredients for Detoxification of Honey-Fired Licorice (ZGC) Based on the Metabolic Profile by UPLC-Q-TOF-MS.

Licorice is well known for its ability to reduce the toxicity of the whole prescription in traditional Chinese medicine theory. However, honey-fired licorice (ZGC for short), which is made of licorice after being stir-fried with honey water, is more commonly used for clinical practice. The metabolism in vivo and detoxification-related compounds of ZGC have not been fully elucidated. In this work, the chemical constituents in ZGC and its metabolic profile in rats were both identified by high ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The network pharmacology was applied to predict the potential detoxifying ingredients of ZGC. As a result, a total of 115 chemical compounds were identified or tentatively characterized in ZGC aqueous extract, and 232 xenobiotics (70 prototypes and 162 metabolites) were identified in serum, heart, liver, kidneys, feces, and urine. Furthermore, 41 compounds absorbed in serum, heart, liver, and kidneys were employed for exploring the detoxification of ZGC by network pharmacology. Ultimately, 13 compounds (five prototypes including P5, P24, P30, P41 and P44, and 8 phase Ⅰ metabolites including M23, M47, M53, M93, M100, M106, M118, and M134) and nine targets were anticipated to be potential mediums regulating detoxification actions. The network pharmacology analysis had shown that the ZGC could detoxify mainly through regulating the related targets of cytochrome P450 and glutathione. In summary, this study would help reveal potential active ingredients in vivo for detoxification of ZGC and provided practical evidence for explaining the theory of traditional Chinese medicine with modern technology.

Comprehensive Identification of Short and Medium-Sized Peptides from Pixian Broad Bean Paste Protein Hydrolysates Using UPLC-Q-TOF-MS and UHPLC-Q Exactive HF-X.

Pixian broad bean paste (PBBP) is an indispensable food widely used in many East Asian countries, yet the knowledge about bioactive peptides released from parent proteins by enzymatic hydrolysis is limited. A total of 5867 low-molecular weight peptides were identified in the highly bioactive subfractions of the PBBP alcalase hydrolysates using traditional and peptidomics approaches. 19 short peptides (3-5 amino acids) were identified by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry, including 5 tripeptides, 8 tetrapeptides, and 6 pentapeptides. 5848 medium-sized peptides (6-10 amino acids) were characterized using the peptidomics approach, including 1484 hexapeptides, 1217 heptapeptides, 1634 octapeptides, 927 nonapeptides, and 586 decapeptides. The comprehensive method can be used for the investigation of bioactive peptides in complex food matrices.

Clinical metabolomic analysis of Danlou tablets with antioxidant effects for treating stable angina pectoris

The Danlou tablets (DLT) is a patented Chinese medicine that can effectively ameliorate coronary heart disease– and angina pectoris–related chest congestion and pain. However, the mechanism underlying the therapeutic effects of DLT in the context of stable angina pectoris (SAP) has not been clearly elucidated. In this study, ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was used to analyze serum samples from patients with SAP before and after DLT administration. The results of metabolomic analyses were verified biologically, and the mechanisms of action of DLT with respect to treating SAP were elucidated. Nineteen biomarkers were identified. Among these biomarkers, the levels of 15 reverted to those corresponding to a healthy state following DLT treatment. The main metabolic pathways associated with the functions of DLT in SAP were energy metabolism, purine metabolism, glycerophospholipid metabolism and amino acid metabolism, all of which are related to oxidative stress. Biological verification revealed that DLT decreased the expression of the oxidative stress indicators, xanthine oxidase (XOD) and malondialdehyde (MDA), and increased heme oxygenase-1 (HO-1) expression and superoxide dismutase (SOD) activity. Taken together, we revealed that DLT effectively ameliorates SAP by adjusting the oxidative stress status. This study provided an objective index for evaluating the efficacy of DLT for treating SAP.

Differentiating ultra-high temperature milk and reconstituted milk using an untargeted peptidomic approach with chemometrics

An untargeted peptide profiling based on ultra-performance liquid chromatography quadrupole time-of flight mass spectrometry with chemometrics was performed to differentiate ultra-high temperature processed milk and reconstituted milk. Thirty-three marker peptides were identified, primarily released from the C- or N-terminal of β-casein and αs1-casein. These peptides were produced by heating and protease hydrolysis. Additional heating and storage time experiments showed that the level of 18 marker peptides increased with heat load and storage time, whereas 15 peptides were solely influenced by heat load. The peptides from β-casein showed higher sensitivity to thermal stress compared to those from αs1-casein. Additionally, eight modified peptides of casein were identified as indicators of milk thermal processing. The identified marker peptides can distinguish ultra-high temperature processed milk and reconstituted milk, and are suitable for monitoring heating processes and storage of milk.

Buyang Huanwu Decoction Ameliorates Damage of Erectile Tissue and Function Following Bilateral Cavernous Nerve Injury.

To verify the effect of Buyang Huanwu Decoction (BHD) in ameliorating erectile dysfunction (ED) after radical prostatectomy (RP). The composition of BHD was verified by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS) analysis. Bilateral cavernous nerve crush injury (BCNI) in rats was used to mimic the neurovascular injury occurring after RP. By the envelope method, forty rats were randomly divided into 4 groups as follows: sham (cavernous nerves exposed only), model (BCNI), low-dosage BHD [LBHD, 12.8 g/(kg·d)], and high-dosage BHD [HBHD, 51.2 g/(kg·d)] groups, 10 rats in each group, feeding for 3 weeks respectively. Erectile function was evaluated by measuring intracavernosal pressure (ICP). Changes in the histopathology of corpus cavernosum (CC) were examined by hematoxylin-eosin staining. Meanwhile, the fibrosis of CC was measured by Masson's trichrome staining and Western blot was used to detect the expressions of collagen I, transforming growth factor beta 1 (TGF- β 1) and α-smooth muscle actin (α-SMA). Apoptosis index was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) and Western blot for determining the expressions of B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X (Bax). The oxidative stress in the CC were assessed by the superoxide dismutase (SOD), malondialdehyde (MDA) and reactive oxygen species (ROS) levels. The proteins expression of c-Jun N-terminal kinase (JNK) and c-Jun were detected by Western blot. In addition, the expression of α-SMA and p-c-Jun in the CC was observed by double immunofluorescence staining. The UPLC-QTOF-MS/MS analysis showed that BHD contained calycosin-7-O- β -D-glucoside, ononin, calycosin and formononetin. Compared with the model group, LBHD and HBHD treatment improved the ICP and the circumference, area, and weight of CC (P<0.05 or P<0.01). Furthermore, LBHD and HBHD treatments increased CC smooth muscle content and decreased apoptosis index (P<0.05 or P<0.01). LBHD and HBHD also elevated SOD and expression level of α -SMA and Bcl-2, and reduced MDA and ROS levels, as well as expression of TGF- β 1, collagen I, Bax, p-c-JNK, p-JNK in the CC compared with the model group (P<0.05 or P<0.01). The double immunofluorescence staining showed that the fluorescence degree of p-c-Jun in both LBHD and HBHD treatment groups was significantly reduced, whereas the α -SMA expression increased (P<0.05 or P<0.01). BHD can improve ED of rats with BCNI, which is related to inhibiting fibrosis, apoptosis, and oxidative stress of CC. The ROS/JNK/c-Jun signaling pathway may play an important role in the process.

Isolation and identification of bitter-tasting peptides in Shaoxing rice wine using ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry combined with taste orientation strategy

The content of peptides in Shaoxing rice wine is high; however, their potential taste properties have not yet been studied. To isolate and identify bitter-tasting peptides in a popular Chinese rice wine, Shaoxing rice wine, a series of purification steps including ultrafiltration and gel filtration chromatography (GFC), combined with a taste orientation strategy, were used. According to the evaluation results of taste dilution analysis, a bitter peptide with the highest taste efficacy was isolated, and its purity was verified by high-performance liquid chromatography (HPLC). The amino acid sequence of peptide components was analyzed by ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS). The bitter peptide Leu-Pro-Thr-Leu (LPTL) was successfully identified from Shaoxing rice wine. Then, taste addition experiments highlighted the bitter impacts of LPTL on sensory descriptors in Shaoxing rice wine. The LPTL peptide can contribute to bitterness in Shaoxing rice wine.

Individualized Dosage of Tacrolimus for Renal Transplantation Patients Based on Pharmacometabonomics.

The clinical pharmacodynamics of tacrolimus in renal transplant patients has significant interindividual variability. T lymphocytes were selected to study the pharmacodynamic response of tacrolimus, which was significantly correlated with renal function and the outcome of renal transplant patients. Ultra-performance liquid chromatography–quadrupole time-of-flight mass spectroscopy (UPLC/Q-TOF-MS) was performed to obtain the metabolic profiles of 109 renal transplant patients. A partial least squares (PLS) model was constructed to screen potential biomarkers that could predict the efficacy of tacrolimus. Multinomial logistic regression analysis established a bridge that could quantify the relationship between the efficacy of tacrolimus and biomarkers. The results showed a good correlation between endogenous molecules and the efficacy of tacrolimus. Metabolites such as serum creatinine, mesobilirubinogen, L-isoleucine, 5-methoxyindoleacetate, eicosapentaenoic acid, N2-succinoylarginine, tryptophyl-arginine, and butyric acid were indicated as candidate biomarkers. In addition, the key biomarkers could correctly predict the efficacy of tacrolimus with an accuracy of 82.5%. Finally, we explored the mechanism of individual variation by pathway analysis, which showed that amino acid metabolism was significantly related to the efficacy of tacrolimus. Moreover, orthogonal partial least squares discriminant analysis (OPLS-DA) showed that there was no difference in key metabolites among different pharmacodynamic groups at 1 month and 3 months after dose adjustment, suggesting that pharmacometabonomics is a useful tool to predict individual differences in pharmacodynamics and thus to facilitate individualized drug therapy.

Integrated Serum Pharmacochemistry and Network Pharmacology Approach to Explore the Effective Components and Potential Mechanisms of Menispermi Rhizoma Against Myocardial Ischemia.

Background: Myocardial ischemia (MI) is a leading cause of death worldwide. Menispermi Rhizoma is a traditional Chinese medicine that exerts a variety of beneficial pharmacological activities in many diseases, including MI. Purpose: Serum pharmacochemistry and network pharmacology were used to explore the material basis and mechanism of action of Menispermi Rhizoma against MI. Methods: The absorbed components of Menispermi Rhizoma in rat plasma were analyzed by ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF/MS). The key components, targets, pathways, and interrelated information were obtained by network pharmacology. The potential effective components of Menispermi Rhizoma against MI were screened by methyl-thiazolyl-tetrazolium (MTT) assay, and the cardioprotective effect and mechanism of active components were verified by Western blotting and molecular docking. Results: In total, 25 absorbed components of Menispermi Rhizoma in plasma were identified. Network pharmacology revealed 81 major targets of Menispermi Rhizoma against MI, mainly involving the regulation of the PI3K/AKT and MAPK pathways. In vitro validation of H9c2 cells revealed that acutumine, daurisoline, dauricoside, and 6-O-demethylmenisporphine are the main bioactive components of Menispermi Rhizoma. The levels of lactate dehydrogenase, creatine kinase, and malondialdehyde (MDA) were significantly decreased by four alkaloids, whereas the activities of superoxide dismutase (SOD) and glutathione (GSH) were significantly increased. Four alkaloids effectively protected H9c2 cells against OGD-induced apoptosis by Hoechst/PI staining and flow cytometry assay. Western blotting results showed that the four alkaloids upregulated the expression ratio of Bcl-2/Bax and downregulated the expression levels of Cyt-C and cleaved caspase 3, which further supported the anti-cardiomyocyte apoptosis and antioxidative stress effect of Menispermi Rhizoma. Molecular docking confirmed that the four compounds were capable of binding to AKT1, MAPK1, EGFR, CASP3, and MAPK8 proteins, suggesting the protective effect of Menispermi Rhizoma on MI via PI3K/AKT, MAPK, and apoptosis pathways. Conclusion: Menispermi Rhizoma exerted cardioprotective effects through the effect characteristics: multiple-ingredient, multi-target, and multi-pathway. This research provided a reference for further mechanistic research on wider applications of Menispermi Rhizoma for MI treatment.

Application of metabolomics analysis to aid in understanding the pathogenicity of different lineages and different serotypes of Listeria monocytogenes

Listeria monocytogenes is a foodborne pathogen with high mortality in young children, elderly persons, pregnant women, and immune-compromised individuals. Most human listeriosis cases are associated with four serotypes (1/2a, 1/2b, 1/2c and 4b) within lineages I and II. The intracellular metabolic changes in L. monocytogenes from different lineages and serotypes remain unclear. Here, a non-targeted metabolomics strategy, based on ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) coupled with chemometrics was performed to analyze the metabolic fluctuations in the lineages and serotypes of L. monocytogenes. In the aggregate, 93 L. monocytogenes isolates (two lineages and four serotypes) were investigated, and a total of 48 differential biomarker metabolites were identified. Twenty differential metabolites were observed at the lineage level, while the others were found at the serotype level. At the lineage level, significantly lower contents (fold change [FC] = 3.60) of γ-aminobutyric acid (GABA) was found in lineage II strains, as compared to lineage I strains. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways enrichment analysis showed that different substances between lineages I and II participated in multiple metabolic pathways, among which pyrimidine metabolism, purine metabolism, and aminoacyl-tRNA biosynthesis were significantly enriched. The major significantly upregulated (P < 0.05) metabolites, including uridine 5′-monophosphate, uridine, guanosine monophosphate, and guanosine, in lineage II strains affect the synthesis of DNA and RNA. At the serotype level, the upregulation of urea precursor substances produced by the purine metabolism pathway in serotype 4b strains may be one reason for the increased virulence of serotype 4b strains, compared to other serotype strains. In addition, the upregulation of pyridoxine in the vitamin B6 pathway in 4b strains was also observed. The differential metabolic pathway of 1/2a and 1/2c strains is nucleotide metabolism, which could lead to differences in the biosynthesis of DNA and RNA of the strain and further affect the growth and proliferation of strains.

Study of Chemical Compositions and Anticancer Effects of Paris polyphylla var. Chinensis Leaves.

Paris polyphylla var. chinensis (Franch.) Hara is a perennial herb belonging to the Trilliaceae family. Ultraperformance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC/Q–TOF MS) was used to detect the composition of different fractions of Paris polyphylla var. chinensis leaves. Meanwhile, the extracts of different fractions were evaluated for their cytotoxic activities against four selected human cancer cell lines and one human normal epithelial cell line based on the MTT assay method. Multivariate statistical analysis was performed to screen differential compounds and to analyze the distributions between different fractions. Finally, more than 60 compounds were obtained and identified from the different fractions of Paris polyphylla var. chinensis leaves, and the chloroform and n−butanol extracts showed significant cytotoxic effects on these four cancer cells. Several compounds were preliminarily identified from different fractions, including 36 steroidal saponins, 11 flavonoids, 10 ceramides, 8 lipids, 6 organic acids, and 8 other compounds. Various compounds were screened out as different chemical components of different fractions, which were considered as a potential substance basis for the cytotoxicity of Paris polyphylla var. chinensis leaves.

Metabolomics study on liver of db/db mice treated with curcumin using UPLC-Q-TOF-MS

A metabolomics method based on ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was applied to study the metabolic changes of liver in db/db mice administered with curcumin. After one week of acclimating to feeding, 20 db/db mice were randomly divided into two groups: curcumin non-treatment group and curcumin treatment group. After eight weeks of treatment, plasma and liver were collected for biochemical analysis and metabolomics analysis, as well as liver oxidative stress and histopathology examination. Serum biochemical indicators such as blood glucose, triglycerides, fasting insulin, and aspartate aminotransferase decreased significantly in the curcumin treatment group compared to the curcumin non-treatment group, whereas hepatic pathological levels and antioxidation levels improved. There were several different potential biomarkers in two groups, including sphingomyelin (SM) (d18:0/20:0), eicosapentaenoic acid (EPA), docosapentaenoic acid (22n-6) (DPAn-6), arachidonic acid (AA), dihomo-gamma- linolenic acid (DGLA), leukotriene C4 (LTC4), lysophosphatidylethanolamine (LysoPE) (16:1(9Z)/0:0), LysoPE (18:1(9Z)/0:0), LysoPE (0:0/18:0), LysoPE (0:0/20:1(11Z)), LysoPE (20:1(11Z)/0:0), 3-sulfinoalanine, alpha-tocotrienol (α-T3) and pantetheine 4′-phosphate (4′-PP). The mechanism might be related to biosynthesis of unsaturated fatty acids, arachidonic acid metabolism, phospholipid metabolism, and taurine and hypotaurine metabolism. The effects of curcumin on type 2 diabetes mellitus (T2DM) include antioxidant, delaying development of T2DM, preventing β-cell death, decreasing insulin resistance, alleviating hepatic damage, and improving metabolic disturbances. Our results provide novel insights and ideas for prevention and treatment of curcumin on T2DM and its complications.