Accurate and sensitive quantitation of glucose and glucose phosphates derived from storage carbohydrates by mass spectrometry

Lyndsay E.A. Young, Corey O. Brizzee, Jessica K.A. Macedo, Robert D. Murphy, Christopher J. Contreras, Anna A. DePaoli-Roach, Peter J. Roach, Matthew S. Gentry, Ramon C. Sun

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

The addition of phosphate groups into glycogen modulates its branching pattern and solubility which all impact its accessibility to glycogen interacting enzymes. As glycogen architecture modulates its metabolism, it is essential to accurately evaluate and quantify its phosphate content. Simultaneous direct quantitation of glucose and its phosphate esters requires an assay with high sensitivity and a robust dynamic range. Herein, we describe a highly-sensitive method for the accurate detection of both glycogen-derived glucose and glucose-phosphate esters utilizing gas-chromatography coupled mass spectrometry. Using this method, we observed higher glycogen levels in the liver compared to skeletal muscle, but skeletal muscle contained many more phosphate esters. Importantly, this method can detect femtomole levels of glucose and glucose phosphate esters within an extremely robust dynamic range with excellent accuracy and reproducibility. The method can also be easily adapted for the quantification of plant starch, amylopectin or other biopolymers.

Original languageEnglish
Article number115651
JournalCarbohydrate Polymers
Volume230
DOIs
StatePublished - Feb 15 2020

Bibliographical note

Funding Information:
This study was supported by National Institute of Neurological Disorders and Stroke (R01 N070899-06, P01 NS097197-01), National Science Foundation (MCB-1817414), the University of Kentucky Center for Cancer and Metabolism, National Institute of General Medical Sciences COBRE program (P20 GM121327), American Cancer Society institutional research grant#16-182-28, and St. Baldrick's Foundation, St Baldrick's Scholar (Career Development Award) – 634297, funding from the University of Kentucky Markey Cancer Center, and the University of Kentucky Epilepsy & Brain Metabolism Alliance. This research was also supported by funding from the University of Kentucky Markey Cancer Center, NIH National Center for Advancing Translational SciencesUL1TR001998, the Biostatistics and Bioinformatics Shared Resource Facility of the University of Kentucky Markey Cancer CenterP30CA177558.

Funding Information:
This study was supported by National Institute of Neurological Disorders and Stroke ( R01 N070899-06 , P01 NS097197-01 ), National Science Foundation ( MCB-1817414 ), the University of Kentucky Center for Cancer and Metabolism , National Institute of General Medical Sciences COBRE program ( P20 GM121327 ), American Cancer Society institutional research grant #16-182-28 , and St. Baldrick’s Foundation, St Baldrick’s Scholar (Career Development Award) – 634297, funding from the University of Kentucky Markey Cancer Center , and the University of Kentucky Epilepsy & Brain Metabolism Alliance . This research was also supported by funding from the University of Kentucky Markey Cancer Center, NIH National Center for Advancing Translational Sciences UL1TR001998 , the Biostatistics and Bioinformatics Shared Resource Facility of the University of Kentucky Markey Cancer Center P30CA177558 .

Publisher Copyright:
© 2019 Elsevier Ltd

Keywords

  • GCMS
  • Glucose
  • Glucose phosphate esters
  • Glycogen
  • Lafora disease
  • Laforin
  • Starch

ASJC Scopus subject areas

  • Organic Chemistry
  • Polymers and Plastics
  • Materials Chemistry

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