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 journalArticle

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 (US)
Article number115651
JournalCarbohydrate Polymers
Volume230
DOIs
StatePublished - Feb 15 2020

Fingerprint

Carbohydrates
Mass spectrometry
Glucose
Glycogen
Phosphates
Esters
Muscle
Amylopectins
Amylopectin
Biopolymers
Starch
Metabolism
Gas chromatography
Liver
Assays
Enzymes
Solubility

Keywords

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

ASJC Scopus subject areas

  • Organic Chemistry
  • Polymers and Plastics
  • Materials Chemistry

Cite this

Accurate and sensitive quantitation of glucose and glucose phosphates derived from storage carbohydrates by mass spectrometry. / Young, Lyndsay E.A.; Brizzee, Corey O.; Macedo, Jessica K.A.; Murphy, Robert D.; Contreras, Christopher J.; DePaoli-Roach, Anna A.; Roach, Peter J.; Gentry, Matthew S.; Sun, Ramon C.

In: Carbohydrate Polymers, Vol. 230, 115651, 15.02.2020.

Research output: Contribution to journalArticle

Young, Lyndsay E.A. ; Brizzee, Corey O. ; Macedo, Jessica K.A. ; Murphy, Robert D. ; Contreras, Christopher J. ; DePaoli-Roach, Anna A. ; Roach, Peter J. ; Gentry, Matthew S. ; Sun, Ramon C. / Accurate and sensitive quantitation of glucose and glucose phosphates derived from storage carbohydrates by mass spectrometry. In: Carbohydrate Polymers. 2020 ; Vol. 230.
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AU - Murphy, Robert D.

AU - Contreras, Christopher J.

AU - DePaoli-Roach, Anna A.

AU - Roach, Peter J.

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AU - Sun, Ramon C.

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KW - Lafora disease

KW - Laforin

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