PubMed

Related Articles Optimizing targeted/untargeted metabolomics by automating gas chromatography/mass spectrometry (GC-GC/MS and GC/MS) workflows. J Chromatogr A. 2017 May 08;: Authors: Robbat A, Kfoury N, Baydakov E, Gankin Y Abstract New database building and MS subtraction algorithms have been developed for automated, sequential two-dimensional gas chromatography/mass spectrometry (GC-GC/MS). This paper reports the first use of a database building tool, with full mass spectrum subtraction, that does not rely on high resolution MS data. The software was used to automatically inspect GC-GC/MS data of high elevation tea from Yunnan, China, to build a database of 350 target compounds. The database was then used with spectral deconvolution to identify 285 compounds by GC/MS of the same tea. Targeted analysis of low elevation tea by GC/MS resulted in the detection of 275 compounds. Non-targeted analysis, using MS subtraction, yielded an additional eight metabolites, unique to low elevation tea. PMID: 28533028 [PubMed - as supplied by publisher]

Related Articles Single nucleotide polymorphisms of ABCC2 modulate renal secretion of endogenous organic anions. Biochem Pharmacol. 2017 May 19;: Authors: Muhrez K, Largeau B, Emond P, Frédéric M, Halimi JM, Trouillas P, Barin-LE Guellec C Abstract The ATP-binding cassette family transporter MRP2 (multidrug resistance-associated protein 2), encoded by the ABCC2 gene, is involved in the renal excretion of numerous xenobiotics and it is likely that it also transports many endogenous molecules arising from not only normal essential metabolic processes but also from environmental toxins or food intake. We used a targeted gas chromatography-mass spectrometry metabolomics analysis to study whether endogenous organic anions are differentially excreted in urines of healthy volunteers according to their genotype for three functional single nucleotide polymorphisms (SNPs) in ABCC2. This was the case for 35 of the 108 metabolites analyzed. Eight of them are most likely substrates of MRP2 since they are the most contributive to the difference between carriers of a decreasing function allele vs those carrying an increasing function one. Seven out of 8 metabolites are fatty acids (dodecanoic acid; 3-hydroxypropanoic acid) or metabolites of polyphenols (caffeine; resorcinol; caffeic acid; 2-(3,4-dihydroxyphenyl) acetic acid; and 4-hydroxyhippuric acid). Most of them were structurally similar to a series of substances previously shown to interact with MRP2 function in vitro. Interestingly, coproporphyrin isomer I, a prototypical substrate of MRP2, also belonged to our final list although it was not significantly discriminant on its own. This suggests that the simultaneous measurement of a set of endogenous metabolites in urine, rather than that of unique metabolites, has the potential to provide a phenotypic measure of MRP2 function in vivo. This would represent an innovative tool to study the variability of the transport activity of MRP2 under a physiological or pathological condition, especially in pharmacokinetic studies of its substrates. PMID: 28532626 [PubMed - as supplied by publisher]

Related Articles Improvement of the omega 3 index of healthy subjects does not alter the effects of dietary saturated fats or n-6PUFA on LDL profiles. Metabolism. 2017 Mar;68:11-19 Authors: Dias CB, Amigó N, Wood LG, Mallol R, Correig X, Garg ML Abstract BACKGROUND AND AIMS: Dietary fat composition is known to modulate circulating lipid and lipoprotein levels. Although supplementation with long chain omega-3 polyunsaturated fatty acids (LCn-3PUFA) has been shown to reduce plasma triglyceride levels, the effect of the interactions between LCn-3PUFA and the major dietary fats consumed has not been previously investigated. METHODS: In a randomized controlled parallel design clinical intervention, we examined the effect of diets rich in either saturated fatty acids (SFA) or omega-6 polyunsaturated fatty acids (n-6PUFA) on plasma lipid levels and lipoprotein profiles (lipoprotein size, concentration and distribution in subclasses) in subjects with an adequate omega 3 index. Twenty six healthy subjects went through a four-week pre-supplementation period with LCn-3PUFA and were then randomized to diets rich in either n-6PUFA or SFA both supplemented with LCn-3PUFA. RESULTS: The diet rich in n-6PUFA decreased low density lipoprotein (LDL) particle concentration (-8%, p=0.013) and LDL cholesterol (LDL-C) level (-8%, p=0.021), while the saturated fat rich diet did not affect LDL particle concentration or LDL-C levels significantly. Nevertheless, dietary saturated fatty acids increased LCn-3PUFA in plasma and tissue lipids compared with n-6PUFA, potentially reducing other cardiovascular risk factors such as inflammation and clotting tendency. CONCLUSION: Improvement on the omega 3 index of healthy subjects did not alter the known effects of dietary saturated fats and n-6PUFA on LDL profiles. PMID: 28183443 [PubMed - indexed for MEDLINE]

Related Articles Serum metabolomic profiling of prostate cancer risk in the prostate, lung, colorectal, and ovarian cancer screening trial. Br J Cancer. 2016 Oct 25;115(9):1087-1095 Authors: Huang J, Mondul AM, Weinstein SJ, Koutros S, Derkach A, Karoly E, Sampson JN, Moore SC, Berndt SI, Albanes D Abstract BACKGROUND: Two recent metabolomic analyses found serum lipid, energy, and other metabolites related to aggressive prostate cancer risk up to 20 years prior to diagnosis. METHODS: We conducted a serum metabolomic investigation of prostate cancer risk in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial that included annual serum total prostate-specific antigen measurement and digital rectal examination. This nested study included 380 cases diagnosed post-screening and 380 controls individually matched to cases on age, race, study centre, and blood-collection date (median time to diagnosis, 10 years (range 4.4-17 years)). Sera were analysed on a high-resolution accurate mass platform of ultrahigh-performance liquid and gas chromatography/mass spectroscopy that identified 695 known metabolites. Logistic regression conditioned on the matching factors estimated odds ratios (OR) and 95% confidence intervals of risk associated with an 80th percentile increase in the log-metabolite signal. RESULTS: Twenty-seven metabolites were associated with prostate cancer at P<0.05. Pyroglutamine, gamma-glutamylphenylalanine, phenylpyruvate, N-acetylcitrulline, and stearoylcarnitine showed the strongest metabolite-risk signals (ORs=0.53, 0.51, 0.46, 0.58, and 1.74, respectively; 0.001⩽P⩽0.006). Findings were similar for aggressive disease (peptide chemical class, P=0.03). None of the P-values were below the threshold of Bonferroni correction, however. CONCLUSIONS: A unique metabolomic profile associated with post-screening prostate cancer is identified that differs from that in a previously studied, unscreened population. PMID: 27673363 [PubMed - indexed for MEDLINE]

Related Articles An Integrated Multi-Omic Approach to Assess Radiation Injury on the Host-Microbiome Axis. Radiat Res. 2016 Sep;186(3):219-34 Authors: Goudarzi M, Mak TD, Jacobs JP, Moon BH, Strawn SJ, Braun J, Brenner DJ, Fornace AJ, Li HH Abstract Medical responders to radiological and nuclear disasters currently lack sufficient high-throughput and minimally invasive biodosimetry tools to assess exposure and injury in the affected populations. For this reason, we have focused on developing robust radiation exposure biomarkers in easily accessible biofluids such as urine, serum and feces. While we have previously reported on urine and serum biomarkers, here we assessed perturbations in the fecal metabolome resulting from exposure to external X radiation in vivo. The gastrointestinal (GI) system is of particular importance in radiation biodosimetry due to its constant cell renewal and sensitivity to radiation-induced injury. While the clinical GI symptoms such as pain, bloating, nausea, vomiting and diarrhea are manifested after radiation exposure, no reliable bioindicator has been identified for radiation-induced gastrointestinal injuries. To this end, we focused on determining a fecal metabolomic signature in X-ray irradiated mice. There is overwhelming evidence that the gut microbiota play an essential role in gut homeostasis and overall health. Because the fecal metabolome is tightly correlated with the composition and diversity of the microorganism in the gut, we also performed fecal 16S rRNA sequencing analysis to determine the changes in the microbial composition postirradiation. We used in-house bioinformatics tools to integrate the 16S rRNA sequencing and metabolomic data, and to elucidate the gut integrated ecosystem and its deviations from a stable host-microbiome state that result from irradiation. The 16S rRNA sequencing results indicated that radiation caused remarkable alterations of the microbiome in feces at the family level. Increased abundance of common members of Lactobacillaceae and Staphylococcaceae families, and decreased abundances of Lachnospiraceae, Ruminococcaceae and Clostridiaceae families were found after 5 and 12 Gy irradiation. The metabolomic data revealed statistically significant changes in the microbial-derived products such as pipecolic acid, glutaconic acid, urobilinogen and homogentisic acid. In addition, significant changes were detected in bile acids such as taurocholic acid and 12-ketodeoxycholic acid. These changes may be associated with the observed shifts in the abundance of intestinal microbes, such as R. gnavus , which can transform bile acids. PMID: 27512828 [PubMed - indexed for MEDLINE]

Related Articles Metabolic profiles of cow's blood; a review. J Sci Food Agric. 2016 Oct;96(13):4321-8 Authors: Puppel K, Kuczyńska B Abstract The term 'metabolic profile' refers to the analysis of blood biochemical parameters that are useful to assess and prevent metabolic and nutritional disorders in dairy herds. In the higher standards of milk production, the priority in modern breeding is keeping dairy cows in high lactation and healthy. The proper analysis, as well as control. of their feeding and metabolic status is immensely important for the health condition of the herd. The disproportion between the genetically determined ability for milk production and the limitations in improving the energy value of the ration may be the cause of metabolic disorders. Negative energy balance has a major impact on the body's hormonal balance and organ functions and mostly appears during transition periods: from 3 to 2 weeks prepartum until 2-3 weeks postpartum. The term 'transition' is used to underscore the important physiological, metabolic and nutritional changes occurring in this time. The manner in which these changes occur and how they are diagnosed and detected are extremely important, as they are closely related to clinical and subclinical postpartum diseases, lactation and reproductive performance - factors that significantly shape the profitability of production. Therefore the priority for intensive milk production is prevention of metabolic diseases and other disorders. It is the intent of this review to synthesize and summarize the information currently available on metabolic status and physiological changes in the cow's body that occur during lactation, as well as to discuss the interpretation of the results, which will be a useful diagnostic tool in nutritional evaluations of the dairy herd. © 2016 Society of Chemical Industry. PMID: 27129620 [PubMed - indexed for MEDLINE]

Workflow methodology for rat brain metabolome exploration using NMR, LC-MS and GC-MS analytical platforms. J Pharm Biomed Anal. 2017 May 13;142:270-278 Authors: Diémé B, Lefèvre A, Nadal-Desbarats L, Galineau L, Madji Hounoum B, Montigny F, Blasco H, Andres CR, Emond P, Mavel S Abstract We developed a multi-platform approach for the metabolome exploration of rat brain tissue, using liquid chromatography coupled with mass spectrometry (LC-MS), nuclear magnetic resonance spectroscopy (NMR) and gas-chromatography coupled with mass spectrometry (GC-MS). The critical steps for metabolite exploration of cerebral tissues are tissue lysis and metabolites extraction. We first evaluated the impact of freeze-drying compared to wet tissue metabolites extraction using NMR and LC-MS with a reversed phase liquid chromatography. Then, we compared four metabolite extraction methods Based on the number of metabolites extracted, their intensity and their coefficient of variation (%CV), the most reproducible protocol (one-step extraction with acetonitrile on lyophilized material) was chosen to further evaluate the impact of sample mass on method performance (3, 6, and 9mg were essayed). GC-MS analysis was also investigated by analyzing four different methoximation/silylation derivatization combinations. The optimal analytical protocols were proposed to establish the reliability required to realize untargeted brain tissue metabolomics exploration. The most reliable workflow was then exemplified by analyzing three rat brain regions (cerebellum, frontal and parietal cortices, n=12) by (1)H NMR, LC-MS and GC-MS, allowing their clustering based on their metabolic profiles. We present here an example of development of methodology that should be done before running analysis campaigns. PMID: 28531831 [PubMed - as supplied by publisher]

Livestock metabolomics and the livestock metabolome: A systematic review. PLoS One. 2017;12(5):e0177675 Authors: Goldansaz SA, Guo AC, Sajed T, Steele MA, Plastow GS, Wishart DS Abstract Metabolomics uses advanced analytical chemistry techniques to comprehensively measure large numbers of small molecule metabolites in cells, tissues and biofluids. The ability to rapidly detect and quantify hundreds or even thousands of metabolites within a single sample is helping scientists paint a far more complete picture of system-wide metabolism and biology. Metabolomics is also allowing researchers to focus on measuring the end-products of complex, hard-to-decipher genetic, epigenetic and environmental interactions. As a result, metabolomics has become an increasingly popular "omics" approach to assist with the robust phenotypic characterization of humans, crop plants and model organisms. Indeed, metabolomics is now routinely used in biomedical, nutritional and crop research. It is also being increasingly used in livestock research and livestock monitoring. The purpose of this systematic review is to quantitatively and objectively summarize the current status of livestock metabolomics and to identify emerging trends, preferred technologies and important gaps in the field. In conducting this review we also critically assessed the applications of livestock metabolomics in key areas such as animal health assessment, disease diagnosis, bioproduct characterization and biomarker discovery for highly desirable economic traits (i.e., feed efficiency, growth potential and milk production). A secondary goal of this critical review was to compile data on the known composition of the livestock metabolome (for 5 of the most common livestock species namely cattle, sheep, goats, horses and pigs). These data have been made available through an open access, comprehensive livestock metabolome database (LMDB, available at http://www.lmdb.ca). The LMDB should enable livestock researchers and producers to conduct more targeted metabolomic studies and to identify where further metabolome coverage is needed. PMID: 28531195 [PubMed - in process]

Metabolic phenotyping of various tea (Camellia sinensis L.) cultivars and understanding of their intrinsic metabolism. Food Chem. 2017 Oct 15;233:321-330 Authors: Ji HG, Lee YR, Lee MS, Hwang KH, Kim EH, Park JS, Hong YS Abstract Recently, we selected three tea (Camellia sinensis) cultivars that are rich in taste, epigallocatechin-3-O-gallate (EGCG) and epigallocatechin-3-O-(3-O-methyl)-gallate (EGCG3″Me) and then cultivated them through asexual propagation by cutting in the same region. In the present study, proton nuclear magnetic resonance ((1)H NMR)-based metabolomics was applied to characterize the metabotype and to understand the metabolic mechanism of these tea cultivars including wild type tea. Of the tea leaf metabolite variations, reverse associations of amino acid metabolism with catechin compound metabolism were found in the rich-taste, and EGCG- and EGCG3″Me-rich tea cultivars. Indeed, the metabolism of individual catechin compounds in the EGCG3″Me-rich cultivar differed from those of other tea cultivars. The current study highlights the distinct metabolism of various tea cultivars newly selected for cultivation and the important role of metabolomics in understanding the metabolic mechanism. Thus, comprehensive metabotyping is a useful method to assess and then develop a new plant cultivar. PMID: 28530581 [PubMed - in process]

Associations of Nasopharyngeal Metabolome and Microbiome with Severity Among Infants with Bronchiolitis: A Multi-omic Analysis. Am J Respir Crit Care Med. 2017 May 21;: Authors: Stewart CJ, Mansbach JM, Wong MC, Ajami NJ, Petrosino JF, Camargo CA, Hasegawa K Abstract RATIONALE: Bronchiolitis is the most common lower respiratory infection in infants; however, it remains unclear which infants with bronchiolitis will develop severe illness. In addition, while emerging evidence indicates associations of the upper-airway microbiome with bronchiolitis severity, little is known about the mechanisms linking airway microbes and host response to disease severity. OBJECTIVES: To determine the relations among the nasopharyngeal airway metabolome profiles, microbiome profiles, and severity in infants with bronchiolitis. METHODS: We conducted a multicenter prospective cohort study of infants (age <1 year) hospitalized with bronchiolitis. By applying metabolomic and metagenomic (16S rRNA gene and whole genome shotgun sequencing) approaches to 144 nasopharyngeal airway samples collected within 24 hours of hospitalization, we determined metabolome and microbiome profiles and their association with higher severity, defined by the use of positive pressure ventilation (PPV) - i.e., continuous positive airway pressure and/or intubation. MEASUREMENTS AND MAIN RESULTS: Nasopharyngeal airway metabolome profiles significantly differed by bronchiolitis severity (P<0.001). Among 254 metabolites identified, a panel of 25 metabolites showed high sensitivity (84%) and specificity (86%) in predicting the use of PPV. The intensity of these metabolites was correlated with relative abundance of Streptococcus pneumoniae. In the pathway analysis, sphingolipid metabolism was the most significantly enriched sub-pathway in infants with PPV use compared to those without (P<0.001). Enrichment of sphingolipid metabolites was positively correlated with the relative abundance of S. pneumoniae. CONCLUSIONS: Our multi-omic analyses demonstrate not only the ability of the metabolomics approach to determine bronchiolitis severity, but also the interrelation between the microbiome and host in bronchiolitis pathobiology. PMID: 28530140 [PubMed - as supplied by publisher]

Related Articles Comparative metabolomic analysis of HPAC cells following the acquisition of erlotinib resistance. Oncol Lett. 2017 May;13(5):3437-3444 Authors: Lee S, Jang WJ, Choi B, Joo SH, Jeong CH Abstract Pancreatic cancer is one of the most lethal types of cancer, due to difficulty in early detection and the limited efficacy of available treatments. Erlotinib is used to inhibit the epidermal growth factor receptor for the treatment of pancreatic cancer; however, erlotinib resistance is a major issue and the mechanisms underlying the development of erlotinib resistance remain unclear. To better understand the alterations in tumor metabolism by acquired resistance to erlotinib, an erlotinib-resistant pancreatic cancer cell line (HPAC-ER) was established, followed by a comparison of the metabolic characteristics between these cells and their erlotinib-sensitive parental cells (HPAC). This comparison was accomplished through mass spectrometry-based targeted metabolic profiling. Five metabolite groups (acylcarnitines, amino acids and biogenic amines, glycerophospholipids, sphingolipids and monosaccharides) were semi-quantified and compared statistically. These results revealed significant differences between the two groups of cells. A significant increase in the level of short-chain acylcarnitines and selected lysophosphatidylcholines, and a significant decrease in the level of acyl-alkyl-phosphatidylcholines and one sphingolipid, were observed in the HPAC-ER cells compared with the HPAC cells. The metabolic changes observed in the present study support the theory that there are increased metabolic demands in erlotinib-resistant cancer, reflecting the changes in acetyl-CoA-associated and choline phospholipid metabolism. These findings will aid in elucidating the changes that occur in pancreatic cancer metabolism through the acquired resistance to erlotinib, and in the identification of biomarkers for the early detection of pancreatic cancer. PMID: 28529573 [PubMed - in process]

Related Articles Anticancer effects of the microbiome and its products. Nat Rev Microbiol. 2017 May 22;: Authors: Zitvogel L, Daillère R, Roberti MP, Routy B, Kroemer G Abstract The human gut microbiome modulates many host processes, including metabolism, inflammation, and immune and cellular responses. It is becoming increasingly apparent that the microbiome can also influence the development of cancer. In preclinical models, the host response to cancer treatment has been improved by modulating the gut microbiome; this is known to have an altered composition in many diseases, including cancer. In addition, cancer treatment with microbial agents or their products has the potential to shrink tumours. However, the microbiome could also negatively influence cancer prognosis through the production of potentially oncogenic toxins and metabolites by bacteria. Thus, future antineoplastic treatments could combine the modulation of the microbiome and its products with immunotherapeutics and more conventional approaches that directly target malignant cells. PMID: 28529325 [PubMed - as supplied by publisher]

Related Articles Pharmacological modulation of autophagy: therapeutic potential and persisting obstacles. Nat Rev Drug Discov. 2017 May 19;: Authors: Galluzzi L, Bravo-San Pedro JM, Levine B, Green DR, Kroemer G Abstract Autophagy is central to the maintenance of organismal homeostasis in both physiological and pathological situations. Accordingly, alterations in autophagy have been linked to clinically relevant conditions as diverse as cancer, neurodegeneration and cardiac disorders. Throughout the past decade, autophagy has attracted considerable attention as a target for the development of novel therapeutics. However, such efforts have not yet generated clinically viable interventions. In this Review, we discuss the therapeutic potential of autophagy modulators, analyse the obstacles that have limited their development and propose strategies that may unlock the full therapeutic potential of autophagy modulation in the clinic. PMID: 28529316 [PubMed - as supplied by publisher]

Related Articles Metabolic insights into the cold survival strategy and overwintering of the common cutworm, Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae). J Insect Physiol. 2017 May 18;: Authors: Zhu W, Zhang H, Meng Q, Wang M, Zhou G, Li X, Wang H, Miao L, Qin Q, Zhang J Abstract The common cutworm, Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae), is a destructive pest in Asia. Although overwintering in the field has not been reported for this species, their larvae are capable of long-term survival in fluctuating temperatures, i.e., 5°C (12 h) plus 13°C (12 h), if food is available. With an increase in climate change due to global warming and the widespread use of greenhouses, further understanding of their cold survival strategy is needed to predict and control their population in the future. In this study, metabolomics was performed to analyze the metabolic features of S. litura larvae exposed to two typical low temperatures: 15°C and 4°C, at which the development, locomotion and feeding activities are maintained or halted, respectively. The results showed that the strategies that regulate lipid and amino acid metabolism were similar at 15°C and 4°C. Cold exposure induced a metabolic shift of energy from carbohydrate to lipid and decreased free amino acids level. Biosynthesis likely contributed to the decrease in amino acids levels even at 4°C, a non-feeding temperature, suggesting an insufficient suppression of anabolism. This explains why food and high temperature pulses are necessary for their long-term cold survival. Glycometabolism was different between 15°C and 4°C. Carbohydrates were used rapidly at 15°C, while trehalose accumulated at 4°C. Interestingly, abundant trehalose and serine are prominent features of Spodoptera exigua larvae, an overwintering species, when compared to S. litura larvae. Exposure to 4°C also induced up-regulation of carbohydrase and protease in the guts of S. litura. Therefore, it is likely that concurrence of food supplement and fluctuating temperatures could facilitate the cold survival of S. litura larvae. We also found that exposure to 4°C could activate the mevalonate pathway in S. litura larvae, which might be related to glycometabolism at 4°C. Overall, our study describes systematically the responses of a cold susceptible insect, S. litura, to low temperatures and explains how fluctuating temperatures facilitate their long-term cold survival indicating the possibility for overwintering of S. litura larvae with global warming and agricultural reforms. PMID: 28529155 [PubMed - as supplied by publisher]

Related Articles Proteomics survey of Solanaceae family: Current status and challenges ahead. J Proteomics. 2017 May 18;: Authors: Ghatak A, Chaturvedi P, Paul P, Agrawal GK, Rakwal R, Kim ST, Weckwerth W, Gupta R Abstract Solanaceae is one of the major economically important families of higher plants and has played a central role in human nutrition since the dawn of human civilization. Therefore, researchers have always been interested in understanding the complex behavior of Solanaceae members to identify key transcripts, proteins or metabolites, which are potentially associated with major traits. Proteomics studies have contributed significantly to understanding the physiology of Solanaceae members. A compilation of all the published reports showed that both gel-based (75%) and gel-free (25%) proteomic technologies have been utilized to establish the proteomes of different tissues, organs, and organelles under normal and adverse environmental conditions. Among the Solanaceae members, most of the research has been focused on tomato (42%) followed by potato (28%) and tobacco (20%), owing to their economic importance. This review comprehensively covers the progress made so far in the field of Solanaceae proteomics including novel methods developed to isolate the proteins from different tissues. Moreover, key proteins presented in this review can serve as a resource to select potential targets for crop improvement. We envisage that information presented in this review would enable us to design the stress tolerant plants with enhanced yields. BIOLOGICAL SIGNIFICANCE: Solanaceae is one of the major economically important families of higher plants and has played a central role in human nutrition since the dawn of human civilization. Therefore, researchers have always been interested in understanding the complex behavior of Solanaceae members to identify key players associated with major traits. Proteomics studies have contributed significantly to understanding the physiology of Solanaceae members. This review comprehensively covers the progress made so far in the field of Solanaceae proteomics including novel methods developed to isolate the proteins from different tissues. Moreover, key proteins presented in this review can serve as a resource to select potential targets for crop improvement. We envisage that information presented in this review would enable us to design the stress tolerant plants with enhanced yields. PMID: 28528990 [PubMed - as supplied by publisher]

Related Articles Performance of a High-Pressure Liquid Chromatography-Ion Mobility-Mass Spectrometry System for Metabolic Profiling. Anal Chem. 2017 May 22;: Authors: Zhang X, Kew K, Reisdorph R, Sartain M, Powell R, Armstrong ML, Quinn K, Cruickshank-Quinn C, Walmsley S, Bokatzian SS, Darland E, Rain M, Imatani K, Reisdorph N Abstract A commercial liquid chromatography/drift tube ion mobility-mass spectrometer (LC/IM-MS) was evaluated for its utility in global metabolomics analysis. Performance was assessed using twelve targeted metabolite standards where the limit of detection (LOD), linear dynamic range, resolving power, and collision cross section (Ω), is reported for each standard. Data was collected in three different instrument operation modes: flow injection analysis with IM-MS (FIA/IM-MS), LC/MS and LC/IM-MS. Metabolomics analyses of human plasma and HaCaT cells were used to compare the above three operation modes. LC/MS provides linearity in response, data processing automation, improved limits of detection, and ease of use. Advantages of LC/IM-MS and FIA/IM-MS include the ability to develop mobility-mass trend lines for structurally similar bio-molecules, increased peak capacity, reduction of chemical/matrix noise, improvement in signal-to-noise, and separations of isobar/isomer compounds that are not resolved by LC. We further tested the feasibility of incorporating IM-MS into conventional LC/MS metabolomics workflows. In general, the addition of ion mobility dimension has increased the separation of compounds in complex biological matrices and has the potential to largely improve the throughput of metabolomics analysis. PMID: 28528542 [PubMed - as supplied by publisher]

Related Articles Lens metabolomic profiling as a tool to understand cataractogenesis in Atlantic salmon and rainbow trout reared at optimum and high temperature. PLoS One. 2017;12(4):e0175491 Authors: Remø SC, Hevrøy EM, Breck O, Olsvik PA, Waagbø R Abstract Periods of high or fluctuating seawater temperatures result in several physiological challenges for farmed salmonids, including an increased prevalence and severity of cataracts. The aim of the present study was to compare cataractogenesis in Atlantic salmon (Salmo salar L.) and rainbow trout (Oncorhynchus mykiss) reared at two temperatures, and investigate whether temperature influences lens metabolism and cataract development. Atlantic salmon (101±2 g) and rainbow trout (125±3 g) were reared in seawater at either 13°C (optimum for growth) or 19°C during the 35 days experiment (n = 4 tanks for each treatment). At the end of the experiment, the prevalence of cataracts was nearly 100% for Atlantic salmon compared to ~50% for rainbow trout, irrespective of temperature. The severity of the cataracts, as evaluated by slit-lamp inspection of the lens, was almost three fold higher in Atlantic salmon compared to rainbow trout. The global metabolic profile revealed differences in lens composition and metabolism between the two species, which may explain the observed differences in cataract susceptibility between the species. The largest differences were seen in the metabolism of amino acids, especially the histidine metabolism, and this was confirmed by a separate quantitative analysis. The global metabolic profile showed temperature dependent differences in the lens carbohydrate metabolism, osmoregulation and redox homeostasis. The results from the present study give new insight in cataractogenesis in Atlantic salmon and rainbow trout reared at high temperature, in addition to identifying metabolic markers for cataract development. PMID: 28419112 [PubMed - indexed for MEDLINE]

Related Articles Generic method for the absolute quantification of glutathione S-conjugates: Application to the conjugates of acetaminophen, clozapine and diclofenac. J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Mar 01;1046:185-194 Authors: den Braver MW, Vermeulen NP, Commandeur JN Abstract Modification of cellular macromolecules by reactive drug metabolites is considered to play an important role in the initiation of tissue injury by many drugs. Detection and identification of reactive intermediates is often performed by analyzing the conjugates formed after trapping by glutathione (GSH). Although sensitivity of modern mass spectrometrical methods is extremely high, absolute quantification of GSH-conjugates is critically dependent on the availability of authentic references. Although (1)H NMR is currently the method of choice for quantification of metabolites formed biosynthetically, its intrinsically low sensitivity can be a limiting factor in quantification of GSH-conjugates which generally are formed at low levels. In the present study, a simple but sensitive and generic method for absolute quantification of GSH-conjugates is presented. The method is based on quantitative alkaline hydrolysis of GSH-conjugates and subsequent quantification of glutamic acid and glycine by HPLC after precolumn derivatization with o-phthaldialdehyde/N-acetylcysteine (OPA/NAC). Because of the lower stability of the glycine OPA/NAC-derivate, quantification of the glutamic acid OPA/NAC-derivate appeared most suitable for quantification of GSH-conjugates. The novel method was used to quantify the concentrations of GSH-conjugates of diclofenac, clozapine and acetaminophen and quantification was consistent with (1)H NMR, but with a more than 100-fold lower detection limit for absolute quantification. PMID: 28189104 [PubMed - indexed for MEDLINE]

Related Articles 1HNMR-Based Discriminatory Analysis of Eurycoma longifolia from Different Locations and Establishing a Profile for Primary Metabolites Identification and Quassinoids Quantification. Planta Med. 2017 Jan;83(1-02):172-182 Authors: Ebrahimi F, Ibrahim B, Teh CH, Murugaiyah V, Lam CK Abstract Quassinoids, the major secondary metabolites of Eurycoma longifolia roots, improve male fertility. Hence, it is crucial to investigate their quantitative level in E. longifolia extracts. A profile was established to identify the primary metabolites and major quassinoids, and quantify quassinoids using external calibration curves. Furthermore, the metabolic discrimination of E. longifolia roots from different regions was investigated. The (1)H-NMR spectra of the quassinoids, eurycomanone, eurycomanol, 13,21-dihydroeurycomanone, and eurycomanol-2-O-β-D-glycopyranoside were obtained. The (1)H-NMR profiles of E. longifolia root aqueous extracts from Perak (n = 30) were obtained and used to identify primary metabolites and the quassinoids. Selangor, Kedah, Terengganu (n = 5 for each), and Perak samples were checked for metabolic discrimination. Hotelling's T(2) plot was used to check for outliers. Orthogonal partial least-squares discriminant analysis was run to reveal the discriminatory metabolites. Perak samples contained formic, succinic, methylsuccinic, fumaric, lactic, acetic and syringic acids as well as choline, alanine, phenylalanine, tyrosine, α-glucose, eurycomanone, eurycomanol, 13,21-dihydroeurycomanone, and eurycomanol-2-O-β-D-glycopyranoside. The extracts from other locations contained the same metabolites. The limit of quantification values were 1.96 (eurycomanone), 15.62 (eurycomanol), 3.91 (13,21-dihydroeurycomanone), and 31.25 (eurycomanol-2-O-β-D-glycopyranoside) ppm. The Hotelling's T(2) plot revealed no outlier. The orthogonal partial least-squares discriminant analysis model showed that choline, eurycomanol, eurycomanol-2-O-β-D-glycopyranoside, and lactic and succinic acid levels were different among regions. Terengganu and Perak samples contained higher amounts of eurycomanol and eurycomanol-2-O-β-D-glycopyranoside, respectively. The current approach efficiently detected E. longifolia root metabolites, quantified the quassinoids, and discriminated E. longifolia roots from different locations. These findings could be applicable to future research on E. longifolia where the higher content of quassinoids is required. PMID: 27399233 [PubMed - indexed for MEDLINE]

Related Articles Targeted metabolomic profiling of cardioprotective effect of Ginkgo biloba L. extract on myocardial ischemia in rats. Phytomedicine. 2016 Jun 01;23(6):621-31 Authors: Wang Z, Zhang J, Ren T, Dong Z Abstract BACKGROUND: Myocardial ischemia (MI) is one of the highest mortality diseases in the world. It is closely associated with metabolism disorders of endogenous substances. Ginkgo biloba L. extract (GBE) is a popular herbal medicine used for prevention and therapy of MI. But its regulation effect on the metabolism disorders caused by MI remains currently unknown. PURPOSE: Our metabolomic profiling study provided insight into endogenous metabolic disorders of MI and cardioprotective mechanisms of GBE. STUDY DESIGN: The rats were preventive administrated of GBE (200mg/kg, i.g.) for 4 weeks and then subcutaneous injected of isoproterenol to establish MI model. Heart marker enzymes and histopathological examination were adopted to evaluate MI model and effect of GBE. On this base, endogenous metabolites in rat plasma and heart were well profiled using the developed targeted metabolomic profiling platform to comprehensively analyze metabolic pathways and find biomarkers. METHODS: A targeted metabolomic profiling platform was developed and only 100μl biological sample was used to quantify 808 metabolites covering the core network of lipid, energy, amino acid and nucleotide metabolism. Then using this platform, endogenous metabolites of rats undergoing MI model and GBE pre-treatment were well profiled. Orthogonal partial least squares discriminant analysis (OPLS-DA) was used to discriminate between groups and find biomarkers. RESULTS: The metabolomic profiles of MI model rats pre-protected by GBE were significantly different from those of unprotected. 47 metabolites were found as potential biomarkers and indicated MI would lead to disturbed metabolism due to inflammation, oxidative stress and structural damage; while GBE could effectively restore fatty acid, sphingolipid, phosphoglyceride, glyceride, amino acid and energy metabolism, closely related to its antioxidant, PAF antagonist and hypolipidemic properties. CONCLUSION: The cardioprotective effect of GBE can be achieved through the comprehensive regulation of multiple metabolic pathways. PMID: 27161403 [PubMed - indexed for MEDLINE]