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Updated: Aug 31, 2025
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The Extraction of Liver Glycogen Molecules for Glycogen Structure Determination
Published on: February 8, 2022
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A Micro-Scale Analytical Method for Determining Glycogen Turnover by NMR and FTMS
Timothy L Scott1, Juan Zhu2, Teresa A Cassel2
1Center for Environmental and Systems Biochemistry and Markey Cancer Center, University of Kentucky, Lexington, KY 40506, USA.
Metabolites
|August 25, 2022
Summary
This study introduces a new microscale method for accurately quantifying glycogen turnover using stable isotope tracing. The technique enables sensitive analysis of glycogen metabolism in various biological samples, advancing cancer and immunology research.
Area of Science:
- Metabolomics
- Biochemistry
- Analytical Chemistry
Background:
- Glycogen is a crucial energy storage molecule found in most tissues, with its metabolism playing a role in cancer and immune cells.
- Accurate analysis of glycogen turnover is essential for understanding these roles, but current methods are time-consuming and lack quantitative precision for stable isotope enrichment.
- Stable isotope tracing, using labeled glucose precursors, requires reliable quantification of both total and labeled glycogen.
Purpose of the Study:
- To develop a microscale method for quantifying intact and acid-hydrolyzed glycogen.
- To enable stable isotope resolved metabolomics (SIRM) analysis of glycogen turnover.
- To provide a sensitive, quantitative, and convenient method for analyzing glycogen in biological samples.
Main Methods:
- Utilized ultra-high-resolution Fourier transform mass spectrometry (UHR-FTMS) and Nuclear Magnetic Resonance (NMR) spectroscopy.
- Employed a rapid microwave-assisted acid hydrolysis for efficient glycogen digestion.
- Optimized digestion parameters (time, temperature, oxygen purging) for >90% recovery.
Main Results:
- Successfully quantified intact and hydrolyzed glycogen in crude biological extracts.
- Applied the method to track 13C6-glucose and 2H7-glucose incorporation into glycogen in various cell and tissue models, including human lung cells, macrophages, murine liver, and patient-derived xenografts.
- Demonstrated the method's utility in tracing glycogen turnover by measuring labeled intermediates like UDP-Glucose and glucose-1-phosphate.
Conclusions:
- The described microscale method offers a sensitive, quantitative, and convenient approach for analyzing glycogen turnover.
- This technique is applicable to milligram amounts of complex biological materials, advancing research in cancer, immunology, and metabolic studies.
- The method facilitates robust stable isotope tracing for glycogen metabolism in diverse cellular and in vivo models.
