14 research outputs found

    Metabolomics: From Method Development to System Biology

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    “Зізнання авантюриста Фелікса Крулля” Томаса Манна як пародія на велику автобіографію

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    Статтю присвячено розгляду рецепції та відображення “Поезії і правди” Й.-В. Гете в романі Т. Манна “Зізнання авантюриста Фелікса Крулля”. Представлена розвідка продовжує ряд наукових досліджень, присвячених вивченню особливостей рецепції автобіографії Й.-В. Гете, зокрема в німецькомовному літературному просторі. При дослідженні особливостей наявного в аналізованому нами романі пародійного наслідування Й.-В. Гете основна увага зосереджується автором на його стилістичному та тематичному рівнях.Статья посвящена рассмотрению рецепции и отражения “Поэзии и правды” Й.-В. Гете в романе Т. Манна “Признания авантюриста Феликса Крулля”. Представленная статья продолжает ряд научных исследований, посвященных изучению особенностей рецепции автобиографии Й.-В. Гете в частности в немецкоязычном литературном пространстве. При изучении особенностей имеющихся в анализированном нами романе пародийного подражания Й.-В. Гете основное внимание автор сосредотачивает на его стилистическом и тематическом уровнях.The article focuses on the reception of Goethe’s “Poetry and Truth” and its reflection in T. Mann’s novel “The Confession of adventurer Felix Crool”. The given article extends the series of scientific investigations that deal with the peculiarities of Goethe’s reception in German literature. Studying the parody imitation peculiarities of Goethe in the analysed novel, the main attention is paid to the stylistic and theme levels

    Transportomics: screening for substrates of ABC transporters in body fluids using vesicular transport assays

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    The ATP-binding cassette (ABC) genes encode the largest family of transmembrane proteins. ABC transporters translocate a wide variety of substrates across membranes, but their physiological function is often incompletely understood. We describe a new method to study the substrate spectrum of ABC transporters: We incubate extracts of mouse urine with membrane vesicles prepared from Spodoptera frugiperda Sf9 insect cells overproducing an ABC transporter and determine the compounds transported into the vesicles by LC/MS-based metabolomics. We illustrate the power of this simple “transportomics” approach using ABCC2, a protein present at sites of uptake and elimination. We identified many new substrates of ABCC2 in urine. These included glucuronides of plant-derived xenobiotics, a class of compounds to which humans are exposed on a daily basis. Moreover, we show that the excretion of these compounds in vivo depends on ABCC2: compared to wild-type mice, the urinary excretion of several glucuronides was increased up to 20-fold in Abcc2-/- mice. Transportomics has broad applicability, as it is not restricted to urine and can be applied to other ATP-dependent transport proteins as well.—Krumpochova, P., Sapthu, S., Brouwers, J. F., de Haas, M., de Vos, R., Borst, P., van de Wetering, K. Transportomics: screening for substrates of ABC transporters in body fluids using vesicular transport assay

    Rapid quantification of underivatized amino acids in plasma by hydrophilic interaction liquid chromatography (HILIC) coupled with tandem mass-spectrometry

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    Background: Amino acidopathies are a class of inborn errors of metabolism (IEM) that can be diagnosed by analysis of amino acids (AA) in plasma. Current strategies for AA analysis include cation exchange HPLC with post-column ninhydrin derivatization, GC-MS, and LC-MS/MS-related methods. Major drawbacks of the current methods are time-consuming procedures, derivative problems, problems with retention, and MS-sensitivity. The use of hydrophilic interaction liquid chromatography (HILIC) columns is an ideal separation mode for hydrophilic compounds like AA. Here we report a HILIC-method for analysis of 36 underivatized AA in plasma to detect defects in AA metabolism that overcomes the major drawbacks of other methods. Methods: A rapid, sensitive, and specific method was developed for the analysis of AA in plasma without derivatization using HILIC coupled with tandem mass-spectrometry (Xevo TQ, Waters). Results: Excellent separation of 36 AA (24 quantitative/12 qualitative) in plasma was achieved on an Acquity BEH Amide column (2.1×100 mm, 1.7 μm) in a single MS run of 18 min. Plasma of patients with a known IEM in AA metabolism was analyzed and all patients were correctly identified. Conclusion: The reported method analyzes 36 AA in plasma within 18 min and provides baseline separation of isomeric AA such as leucine and isoleucine. No separation was obtained for isoleucine and allo-isoleucine. The method is applicable to study defects in AA metabolism in plasma

    Metabolomics: From Method Development to System Biology

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    Teusink, B. [Promotor]Somsen, G.W. [Promotor]Giera, M.A. [Copromotor

    Amino acid analysis using chromatography–mass spectrometry: An inter platform comparison study

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    The analysis of amino acids has become a central task in many aspects. While amino acid analysis has traditionally mainly been carried out using either gas chromatography (GC) in combination with flame ionization detection or liquid chromatography (LC) with either post-column derivatization using ninhydrin or pre-column derivatization using o-phthalaldehyde, many of today's analysis platforms are based on chromatography in combination with mass spectrometry (MS). While derivatization is mandatory for the GC-based analysis of amino acids, several LC platforms have emerged, particularly in the dawn of targeted metabolite profiling using hydrophilic interaction liquid chromatography (HILIC) coupled to MS, allowing the analysis of underivatized amino acids. Among the numerous analytical platforms available for amino acid analysis today, we here compare three prominent approaches, being GC-MS and LC-MS after amino acid derivatization using chloroformate and HILIC-MS of underivatized amino acids. We compare and discuss practical issues as well as performance characteristics, e.g., the use of 13C-labeled internal standards, of the different platforms and present data on their practical implementation in our laboratory. Finally, we compare the real-life applicability of all three platforms for a complex biological sample. While all three platforms are very-well suited for the analysis of complex biological samples they all show advantages and disadvantages for some analytes as discussed in detail in this manuscript
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