Determination of the Isotopic Enrichment of 13C- and 2H-Labeled Tracers of Glucose Using High-Resolution Mass Spectrometry: Application to Dual- and Triple-Tracer Studies

Martin Trötzmüller, Alexander Triebl, Amra Ajsic, Jürgen Hartler, Harald Köfeler, Werner Regittnig

Publikation: Beitrag in einer FachzeitschriftArtikelForschungBegutachtung

Abstract

Multiple-tracer approaches for investigating glucose metabolism in humans usually involve the administration of stable and radioactive glucose tracers and the subsequent determination of tracer enrichments in sampled blood. When using conventional, low-resolution mass spectrometry (LRMS), the number of spectral interferences rises rapidly with the number of stable tracers employed. Thus, in LRMS, both computational effort and statistical uncertainties associated with the correction for spectral interferences limit the number of stable tracers that can be simultaneously employed (usually two). Here we show that these limitations can be overcome by applying high-resolution mass spectrometry (HRMS). The HRMS method presented is based on the use of an Orbitrap mass spectrometer operated at a mass resolution of 100 000 to allow electrospray-generated ions of the deprotonated glucose molecules to be monitored at their exact masses. The tracer enrichment determination in blood plasma is demonstrated for several triple combinations of 13C- and 2H-labeled glucose tracers (e.g., [1-2H1]-, [6,6-2H2]-, [1,6-13C2]glucose). For each combination it is shown that ions arising from 2H-labeled tracers are completely differentiated from those arising from 13C-labeled tracers, thereby allowing the enrichment of a tracer to be simply calculated from the observed ion intensities using a standard curve with curve parameters unaffected by the presence of other tracers. For each tracer, the HRMS method exhibits low limits of detection and good repeatability in the tested 0.1-15.0% enrichment range. Additionally, due to short sample preparation and analysis times, the method is well-suited for high-throughput determination of multiple glucose tracer enrichments in plasma samples.

Originalspracheenglisch
Seiten (von - bis)12252-12260
Seitenumfang9
FachzeitschriftAnalytical Chemistry
Jahrgang89
Ausgabenummer22
DOIs
PublikationsstatusVeröffentlicht - 21 Nov 2017

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Mass spectrometry
Glucose
Ions
Blood
Plasmas
Mass spectrometers
Metabolism
Throughput
Molecules

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    Determination of the Isotopic Enrichment of 13C- and 2H-Labeled Tracers of Glucose Using High-Resolution Mass Spectrometry : Application to Dual- and Triple-Tracer Studies. / Trötzmüller, Martin; Triebl, Alexander; Ajsic, Amra; Hartler, Jürgen; Köfeler, Harald; Regittnig, Werner.

    in: Analytical Chemistry, Jahrgang 89, Nr. 22, 21.11.2017, S. 12252-12260.

    Publikation: Beitrag in einer FachzeitschriftArtikelForschungBegutachtung

    Trötzmüller, Martin ; Triebl, Alexander ; Ajsic, Amra ; Hartler, Jürgen ; Köfeler, Harald ; Regittnig, Werner. / Determination of the Isotopic Enrichment of 13C- and 2H-Labeled Tracers of Glucose Using High-Resolution Mass Spectrometry : Application to Dual- and Triple-Tracer Studies. in: Analytical Chemistry. 2017 ; Jahrgang 89, Nr. 22. S. 12252-12260.
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    abstract = "Multiple-tracer approaches for investigating glucose metabolism in humans usually involve the administration of stable and radioactive glucose tracers and the subsequent determination of tracer enrichments in sampled blood. When using conventional, low-resolution mass spectrometry (LRMS), the number of spectral interferences rises rapidly with the number of stable tracers employed. Thus, in LRMS, both computational effort and statistical uncertainties associated with the correction for spectral interferences limit the number of stable tracers that can be simultaneously employed (usually two). Here we show that these limitations can be overcome by applying high-resolution mass spectrometry (HRMS). The HRMS method presented is based on the use of an Orbitrap mass spectrometer operated at a mass resolution of 100 000 to allow electrospray-generated ions of the deprotonated glucose molecules to be monitored at their exact masses. The tracer enrichment determination in blood plasma is demonstrated for several triple combinations of 13C- and 2H-labeled glucose tracers (e.g., [1-2H1]-, [6,6-2H2]-, [1,6-13C2]glucose). For each combination it is shown that ions arising from 2H-labeled tracers are completely differentiated from those arising from 13C-labeled tracers, thereby allowing the enrichment of a tracer to be simply calculated from the observed ion intensities using a standard curve with curve parameters unaffected by the presence of other tracers. For each tracer, the HRMS method exhibits low limits of detection and good repeatability in the tested 0.1-15.0{\%} enrichment range. Additionally, due to short sample preparation and analysis times, the method is well-suited for high-throughput determination of multiple glucose tracer enrichments in plasma samples.",
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    T1 - Determination of the Isotopic Enrichment of 13C- and 2H-Labeled Tracers of Glucose Using High-Resolution Mass Spectrometry

    T2 - Application to Dual- and Triple-Tracer Studies

    AU - Trötzmüller, Martin

    AU - Triebl, Alexander

    AU - Ajsic, Amra

    AU - Hartler, Jürgen

    AU - Köfeler, Harald

    AU - Regittnig, Werner

    PY - 2017/11/21

    Y1 - 2017/11/21

    N2 - Multiple-tracer approaches for investigating glucose metabolism in humans usually involve the administration of stable and radioactive glucose tracers and the subsequent determination of tracer enrichments in sampled blood. When using conventional, low-resolution mass spectrometry (LRMS), the number of spectral interferences rises rapidly with the number of stable tracers employed. Thus, in LRMS, both computational effort and statistical uncertainties associated with the correction for spectral interferences limit the number of stable tracers that can be simultaneously employed (usually two). Here we show that these limitations can be overcome by applying high-resolution mass spectrometry (HRMS). The HRMS method presented is based on the use of an Orbitrap mass spectrometer operated at a mass resolution of 100 000 to allow electrospray-generated ions of the deprotonated glucose molecules to be monitored at their exact masses. The tracer enrichment determination in blood plasma is demonstrated for several triple combinations of 13C- and 2H-labeled glucose tracers (e.g., [1-2H1]-, [6,6-2H2]-, [1,6-13C2]glucose). For each combination it is shown that ions arising from 2H-labeled tracers are completely differentiated from those arising from 13C-labeled tracers, thereby allowing the enrichment of a tracer to be simply calculated from the observed ion intensities using a standard curve with curve parameters unaffected by the presence of other tracers. For each tracer, the HRMS method exhibits low limits of detection and good repeatability in the tested 0.1-15.0% enrichment range. Additionally, due to short sample preparation and analysis times, the method is well-suited for high-throughput determination of multiple glucose tracer enrichments in plasma samples.

    AB - Multiple-tracer approaches for investigating glucose metabolism in humans usually involve the administration of stable and radioactive glucose tracers and the subsequent determination of tracer enrichments in sampled blood. When using conventional, low-resolution mass spectrometry (LRMS), the number of spectral interferences rises rapidly with the number of stable tracers employed. Thus, in LRMS, both computational effort and statistical uncertainties associated with the correction for spectral interferences limit the number of stable tracers that can be simultaneously employed (usually two). Here we show that these limitations can be overcome by applying high-resolution mass spectrometry (HRMS). The HRMS method presented is based on the use of an Orbitrap mass spectrometer operated at a mass resolution of 100 000 to allow electrospray-generated ions of the deprotonated glucose molecules to be monitored at their exact masses. The tracer enrichment determination in blood plasma is demonstrated for several triple combinations of 13C- and 2H-labeled glucose tracers (e.g., [1-2H1]-, [6,6-2H2]-, [1,6-13C2]glucose). For each combination it is shown that ions arising from 2H-labeled tracers are completely differentiated from those arising from 13C-labeled tracers, thereby allowing the enrichment of a tracer to be simply calculated from the observed ion intensities using a standard curve with curve parameters unaffected by the presence of other tracers. For each tracer, the HRMS method exhibits low limits of detection and good repeatability in the tested 0.1-15.0% enrichment range. Additionally, due to short sample preparation and analysis times, the method is well-suited for high-throughput determination of multiple glucose tracer enrichments in plasma samples.

    KW - Journal Article

    KW - Chromatography, Liquid

    KW - Reproducibility of Results

    KW - Mass spectrometry

    KW - High-throughput screening

    KW - Software

    KW - Metabolic Networks and Pathways

    KW - Tracer studies

    U2 - 10.1021/acs.analchem.7b03134

    DO - 10.1021/acs.analchem.7b03134

    M3 - Article

    VL - 89

    SP - 12252

    EP - 12260

    JO - Analytical Chemistry

    JF - Analytical Chemistry

    SN - 0003-2700

    IS - 22

    ER -