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

doi:10.1016/j.carbpol.2019.115651

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 journal › Article › peer-review

32 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 language	English
Article number	115651
Journal	Carbohydrate Polymers
Volume	230
DOIs	https://doi.org/10.1016/j.carbpol.2019.115651
State	Published - Feb 15 2020

Bibliographical note

Publisher Copyright:
© 2019 Elsevier Ltd

Funding

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. 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 .

Funders	Funder number
NIH National Center for Advancing Translational SciencesUL1TR001998
NIH-funded University of Kentucky Center for Cancer and Metabolism
University of Kentucky Epilepsy & Brain Metabolism Alliance
National Science Foundation Arctic Social Science Program	MCB-1817414
American Cancer Society-Michigan Cancer Research Fund	16-182-28
National Childhood Cancer Registry – National Cancer Institute	P30CA177558
National Institute of General Medical Sciences DP2GM119177 Sophie Dumont National Institute of General Medical Sciences	P20 GM121327
Institute of Neurological Disorders and Stroke National Advisory Neurological Disorders and Stroke Council	R01 N070899-06, P01 NS097197-01
St. Baldrick's Foundation	634297
National Center for Advancing Translational Sciences (NCATS)	UL1TR001998
University of Kentucky Markey Comprehensive Cancer Center

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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10.1016/j.carbpol.2019.115651

Cite this

Young, L. E. A., Brizzee, C. O., Macedo, J. K. A., Murphy, R. D., Contreras, C. J., DePaoli-Roach, A. A., Roach, P. J., Gentry, M. S., & Sun, R. C. (2020). Accurate and sensitive quantitation of glucose and glucose phosphates derived from storage carbohydrates by mass spectrometry. Carbohydrate Polymers, 230, Article 115651. https://doi.org/10.1016/j.carbpol.2019.115651

@article{2916123173af45988943794290bb76fe,

title = "Accurate and sensitive quantitation of glucose and glucose phosphates derived from storage carbohydrates by mass spectrometry",

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.",

keywords = "GCMS, Glucose, Glucose phosphate esters, Glycogen, Lafora disease, Laforin, Starch",

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

note = "Publisher Copyright: {\textcopyright} 2019 Elsevier Ltd",

year = "2020",

month = feb,

day = "15",

doi = "10.1016/j.carbpol.2019.115651",

language = "English",

volume = "230",

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T1 - Accurate and sensitive quantitation of glucose and glucose phosphates derived from storage carbohydrates by mass spectrometry

AU - Young, Lyndsay E.A.

AU - Brizzee, Corey O.

AU - Macedo, Jessica K.A.

AU - Murphy, Robert D.

AU - Contreras, Christopher J.

AU - DePaoli-Roach, Anna A.

AU - Roach, Peter J.

AU - Gentry, Matthew S.

AU - Sun, Ramon C.

PY - 2020/2/15

Y1 - 2020/2/15

N2 - 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.

AB - 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.

KW - GCMS

KW - Glucose

KW - Glucose phosphate esters

KW - Glycogen

KW - Lafora disease

KW - Laforin

KW - Starch

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SN - 0144-8617

VL - 230

JO - Carbohydrate Polymers

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M1 - 115651

ER -

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

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

Access to Document

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