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

Research output: Contribution to journalArticleResearchpeer-review

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.

LanguageEnglish
Pages12252-12260
Number of pages9
JournalAnalytical Chemistry
Volume89
Issue number22
DOIs
StatusPublished - 21 Nov 2017

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Mass spectrometry
Glucose
Ions
Blood
Plasmas
Mass spectrometers
Metabolism
Throughput
Molecules

Keywords

  • Journal Article
  • Chromatography, Liquid
  • Reproducibility of Results
  • Mass spectrometry
  • High-throughput screening
  • Software
  • Metabolic Networks and Pathways
  • Tracer studies

Fields of Expertise

  • Human- & Biotechnology
  • Information, Communication & Computing

Treatment code (Nähere Zuordnung)

  • Application
  • Basic - Fundamental (Grundlagenforschung)

Cooperations

  • BioTechMed-Graz

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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, Vol. 89, No. 22, 21.11.2017, p. 12252-12260.

Research output: Contribution to journalArticleResearchpeer-review

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 ; Vol. 89, No. 22. pp. 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 - Analytical Chemistry

AU - Trötzmüller,Martin

AU - Triebl,Alexander

AU - Ajsic,Amra

AU - Hartler,Jürgen

AU - Köfeler,Harald

AU - Regittnig,Werner

PY - 2017/11/21

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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

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VL - 89

SP - 12252

EP - 12260

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 22

ER -