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Enthalpy changes are typically tabulated for reactions in which both the reactants and products are at the same conditions. A standard state is a commonly accepted set of conditions used as a reference point for the determination of properties under other different conditions. For chemists, the IUPAC standard state refers to materials under a pressure of 1 bar and solutions at 1 M and does not specify a temperature. Many thermochemical tables list values with a standard state of 1 atm. Because...
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On comparing the reactivity of silver and lead, it is observed that the two ionic species, Ag+ (aq) and Pb2+ (aq), show a difference in their redox reactivity towards copper: the silver ion undergoes spontaneous reduction, while the lead ion does not. This relative redox activity can be easily quantified in electrochemical cells by a property called cell potential. This property is commonly known as cell voltage in electrochemistry, and it is a measure of the energy which accompanies the charge...
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Standardization of PGC-LC-MS-based glycomics for sample specific glycotyping.

Christopher Ashwood1, Brian Pratt2, Brendan X MacLean2

  • 1Department of Molecular Sciences, Macquarie University, Sydney, NSW, Australia. nicki.packer@mq.edu.au and ARC Centre of Excellence for Nanoscale Biophotonics, Macquarie University, Sydney, NSW, Australia and Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI, USA.

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|May 9, 2019
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Summary

This study introduces a method to normalize glycan analysis using porous graphitized carbon chromatography, enabling system-independent retention values for better glycomics discovery and quantitation.

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Area of Science:

  • Glycomics
  • Chromatography
  • Mass Spectrometry

Background:

  • Porous graphitized carbon (PGC) chromatography offers high-resolution glycan separation, crucial for identifying isomers and novel structures.
  • Current PGC applications in glycomics are limited by the lack of system-independent retention values for normalization.
  • Technical variations in retention time and peak area hinder reproducible glycan analysis.

Purpose of the Study:

  • To establish system-independent retention values for PGC-based glycan analysis.
  • To develop an automated normalization method for glycomics studies.
  • To create a comprehensive library of PGC-separated glycan structures.

Main Methods:

  • Utilized hydrolyzed dextran as an internal standard for normalization.
  • Employed Skyline software for post-acquisition data processing.
  • Developed a spectral MS/MS library of a dextran ladder for automated normalization.

Main Results:

  • Assigned system-independent glucose unit (GU) retention values for over 300 PGC-separated glycans.
  • Created predictive models for core-fucosylation and bisecting GlcNAc modifications.
  • Successfully discriminated between cell culture and tissue samples based on N-glycan intensity.

Conclusions:

  • The developed normalization method enhances the reliability and automation of PGC-based glycan identification and quantitation.
  • The GU retention value library and predictive models facilitate glycan structure discovery.
  • This approach supports advanced glycomics research and biomarker discovery.