Liquid Chromatography Quadruple Time-of-flight Mass Spectrometry Research Articles

Metabolomics analysis reveals the mechanisms of the effect of Sijunzi decoction on spleen deficiency syndrome in a rat model

Sijunzi decoction (SJZD) is a traditional Chinese medicine prescription which is used to treat spleen deficiency syndrome. Its mechanism of action was studied in a rat model of reserpine-induced spleen deficiency syndrome with metabolomics assays performed in normal control, model and Sijunzi decoction treated rats by liquid chromatography quadruple-time-of-flight mass spectrometry (LC-Q/TOF-MS). Enzyme assays and histopathology, with multivariate statistical analysis, showed that Sijunzi decoction was an effective intervention in model rats. Principal component analysis (PCA) revealed that the metabolite profiles of the model rats and normal control rats were completely separated, and that the profile of the Sijunzi decoction treated rats was similar to that of normal controls. Eighty potential biomarkers of Sijunzi decoction treatment were screened. The most relevant metabolic pathways were glycerophospholipid metabolism, elongation of fatty acids in mitochondria, synthesis and degradation of ketone bodies, fatty acid metabolism, ether lipid metabolism, synthesis of strontium scaffold, proline, degradation of leucine and isoleucine, and metabolism of methyl butyrate. An interaction network of metabolic pathways associated with potential biomarkers was constructed. Then, three potential target molecules of Sijunzi decoction intervention spleen deficiency syndrome including acetyl-CoA, 3-hydroxy-3-methylglutaryl-CoA and hexanoyl-CoA were screened. This study revealed the mechanism of Sijunzi decoction for the treatment of spleen deficiency syndrome and provided a theoretical basis for the clinical application of Sijunzi decoction.

Anti-Influenza Virus Activity of Adlay Tea Components.

Our previous study showed anti-influenza virus activity in adlay tea prepared from adlay seeds, naked barley seeds, soybean, and cassia seeds. In this study, we evaluated the anti-influenza virus activity of each component of this tea and analyzed their active ingredients. Each component was roasted and extracted in hot water; the extracts were tested for antiviral activity and their mechanisms of action were studied. All the tea components showed antiviral activity against the H1N1 and H3N2 influenza subtypes and against influenza B. The viral stages inhibited by the components were virus adsorption and replication in proliferative process, suggesting that the action mechanisms of the components might differ from those of oseltamivir acid. Of the tea components, soybean showed the strongest activity. Therefore, we analyzed its active ingredients by liquid chromatography quadruple time-of-flight mass spectrometry (LC/qTOF-MS) and daidzein and glycitein were detected as active ingredients. Here, anti-influenza virus action of glycitein was the first report.

Early potential metabolic biomarkers of primary postpartum haemorrhage based on serum metabolomics.

Postpartum hemorrhage (PPH) is the leading cause of maternal death, accounting for 1/4 of maternal deaths worldwide. Determining sensitive biomarkers in the peripheral blood to identify postpartum haemorrhage (PPH) is essential for the early diagnosis and management of PPH. The purpose of this study is to identify predictive serum metabolic biomarkers of PPH. Thirty healthy pregnant women and 30 cases of postpartum hemorrhage were studied for our research. The serum metabolites of all pregnant were detected by liquid chromatography-quadruple time-of-flight mass spectrometry (LC-QTOFMS) and the corresponding biomarkers were identified. 34 significantly altered metabolites in PPH-pre-group were identified. They were mainly involved in fatty acid, and glycerophospholipid metabolism. The LysoPCs, PCs, PGs, PIs were effective biomarkers for identifying PPH. The disturbed signaling pathways, mTOR signaling, acute phase response signaling, AMPK signaling and eNOS signaling pathways might be related to the etiopathogenesis of PPH. Our study provided a valuable attempt to screen early diagnostic markers of PPH and to further understand its pathogenesis.

Identification of Eight Synthetic Cannabinoids, Including 5F-AKB48 in Seized Herbal Products Using DART-TOF-MS and LC-QTOF-MS as Nontargeted Screening Methods.

Synthetic cannabinoids are sprayed onto plant material and smoked for their marijuana-like effects. Clandestine manufacturers modify synthetic cannabinoid structures by creating closely related analogs. Forensic laboratories are tasked with detection of these analog compounds, but targeted analytical methods are often thwarted by the structural modifications. Here, direct analysis in real time coupled to accurate mass time-of-flight mass spectrometry (DART-TOF-MS) in combination with liquid chromatography quadruple time-of-flight mass spectrometry (LC-QTOF-MS) are presented as a screening and nontargeted confirmation method, respectively. Methanol extracts of herbal material were run using both methods. Spectral data from four different herbal products were evaluated by comparing fragmentation pattern, accurate mass and retention time to available reference standards. JWH-018, JWH-019, AM2201, JWH-122, 5F-AKB48, AKB48-N-(4-pentenyl) analog, UR144, and XLR11 were identified in the products. Results demonstrate that DART-TOF-MS affords a useful approach for rapid screening of herbal products for the presence and identification of synthetic cannabinoids.

Comparison of sepsis rats induced by caecal ligation puncture or Staphylococcus aureus using a LC-QTOF-MS metabolomics approach

Sepsis is a whole-body inflammatory response to infection with high mortality and is treated in intensive care units (ICUs). In the present study, to identify metabolic biomarkers that can differentiate sepsis models induced by caecal ligation puncture (CLP) or Staphylococcus aureus (S. aureus), small molecular metabolites in the serum were measured by liquid chromatography quadruple time-of-flight mass spectrometry (LC-QTOF-MS) and analysed using the multivariate statistical analysis (MVA) of partial least square-discrimination analysis (PLS-DA) method. The results demonstrated that the body showed obvious metabolic disorders in the sepsis groups compared with the control group. A total of 8 potential biomarkers were identified in the CLP group, and 10 potential biomarkers were identified in the S. aureus group. These potential biomarkers primarily reflected an energy metabolism disorder, inflammatory response, oxidative stress and tissue damage, which occur during sepsis, and these markers might potentially be used to differentiate CLP from Staphylococcus aureus sepsis.

New potential biomarkers for mesterolone misuse in human urine by liquid chromatography quadrupole time-of-flight mass spectrometry.

In this paper, mesterolone metabolic profiles were investigated carefully. Mesterolone was administered to one healthy male volunteer. Urinary extracts were analyzed by liquid chromatography quadruple time-of-flight mass spectrometry (LC-QTOFMS) for the first time. Liquid-liquid extraction was applied to processing urine samples, and dilute-shoot analyses of intact metabolites were also presented. In LC-QTOFMS analysis, chromatographic peaks for potential metabolites were hunt down by using the theoretical [M-H](-) as target ions in full scan experiment, and their actual deprotonated ions were analyzed in targeted MS/MS mode. Ten metabolites including seven new sulfate and three glucuronide conjugates were found for mesterolone. Because of no useful fragment ion for structural elucidation, gas chromatography-mass spectrometry instrumentation was employed to obtain structural details of the trimethylsilylated phase I metabolite released after solvolysis. Thus, their potential structures were proposed particularly by a combined MS approach. All the metabolites were also evaluated in terms of how long they could be detected, and S1 (1α-methyl-5α-androst-3-one-17β-sulfate) together with S2 (1α-methyl-5α-androst-17-one-3β-sulfate) was detected up to 9 days after oral administration, which could be the new potential biomarkers for mesterolone misuse.

A new potential biomarker for 1-testosterone misuse in human urine by liquid chromatography quadruple time-of-flight mass spectrometry

In this study, 1-testosterone urinary metabolic profiles were investigated by liquid chromatography quadruple time-of-flight mass spectrometry (LC-QTOF-MS) using full scan and targeted MS/MS techniques with accurate mass measurement.

Joint MS‐based platforms for comprehensive comparison of rat plasma and serum metabolic profiling

Metabolomics is a rapidly growing field in the comprehensive understanding of cellular and organism-specific responses associated with perturbations induced by medicines, chemicals and environment. Blood matrices are frequently used in clinical and biological studies. In this study, we compared metabolic profiling between rat plasma and serum using complementary platforms of gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-quadruple time-of-flight-mass spectrometry (LC-QTOF-MS). The sample types that were tested included plasma prepared with K2 EDTA and serum collected using venous blood collection protocols. The results of peak area variation for each detected metabolite/feature in the quality control samples showed a good reproducibility in LC-QTOF-MS and better reproducibility in GC-MS. In GC-MS analysis: (a) 25.8% of the defined metabolites differed serum from plasma profiling (t-test, p < 0.05); and (b) serum possessed higher sensitivity than plasma for its generally higher peak intensity in the metabolic profiling. In LC-QTOF-MS analysis, 13 (in positive ion mode) and seven (in negative ion mode) important metabolites were identified as mainly contributing to the separation between serum and plasma.

Metabolic profiling reveals therapeutic biomarkers of processed Aconitum Carmichaeli Debx in treating hydrocortisone induced Kidney-Yang deficiency syndrome rats

Ethnopharmacological relevanceKidney-Yang Deficiency syndrome (KYDS) is a diagnostic pattern in traditional Chinese medicine (TCM) and clinical data showed that the unbalance in adrenal cortical hormone is the key issue in KYDS patients. The processed Ranunculaceae Aconitum carmichaeli debx (Bai-Fu-Pian in Chinese, BFP) is one of the most commonly used Chinese herbs for treating KYDS. The present study was conducted to explore the therapeutic biomarkers of the BFP in treating hydrocortisone administration induced KYDS rats. Materials and methodsThirty male Sprague-Dawley rats were randomly divided into five groups with six in each group. KYDS in rats was induced by i.p. injection of hydrocortisone at the dose of 10mg/kg per day for 15 days as described previously. The rats with KYDS were administered orally, starting from the day of hydrocortisone administration stopped, with BFP extract at the dose of 0.32g/kg, 0.64g/kg and 1.28g/kg per day respectively for 15 days. The blood samples were collected for the liquid chromatography quadruple time-of-flight mass spectrometry (LC-Q-TOF-MS) test, as well as radioimmunoassay to determine the concentrations of cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP) and adrenocorticotrophic hormone (ACTH). The metabolic responses to BFP administration were investigated by using the principal components analysis (PCA) and orthogonal partial least squares analysis (OPLS). Bioinformatics analyses were performed by using the Ingenuity Pathway Analysis (IPA). Variance analysis and linear regression analysis were used in this study. ResultsThe signs and concentrations of cAMP, cGMP and ACTH in the model rats were similar to those previously described about KYDS rats and BFP treatment can reverse the changes. Seventeen significantly changed metabolites among different groups were identified. Thirteen metabolites were identified in the KYDS rats comparing to healthy rats with nine up-regulated and four down-regulated. After BFP treatment at three dosages, five up-regulated metabolites including phosphate, betaine, (4-hydroxyphenyl) acetaldehyde, 5-hydroxyindol-3-acetic acid and 5'-phosphoribosyl-N-formylglycinamide were dose-dependently reversed. The network analysis with IPA showed that four canonical pathways including superpathway of methionine degradation, purine nucleotides De Novo biosynthesis II, tyrosine synthesis and serotonin receptor signaling involved the therapeutic mechanism of BFP in treating the KYDS rats. ConclusionsFive therapeutic biomarkers (phosphate, betaine, (4-hydroxyphenyl) acetaldehyde, 5-hydroxyindol-3-acetic acid and 5'-phosphoribosyl-N-formylglycinamide) and two corresponding canonical pathways (amino acid metabolism and purine nucleotide metabolism) were identified to be involved in the therapeutic mechanism of BFP treating the KYDS.