TY - JOUR
T1 - A new LC-MS/MS technique for separation of gangliosides using a phenyl-hexyl column
T2 - Systematic separation according to sialic acid class and ceramide subclass
AU - Gobburi, Ashta Lakshmi Prasad
AU - Kipruto, Eric Wekesa
AU - Inman, Denise M.
AU - Anderson, David J.
N1 - Funding Information:
The authors thank Dr. Tariq M. Haqqi for use of the LC-MS/MS instrumentation in his laboratory at Northeast Ohio Medical University, Rootstown, Ohio. This work was conducted with support from the Faculty and Research Development Program and the Dissertation Research Award program at Cleveland State University. This work was also supported by the Neurodegenerative Diseases and Aging Research Focus Area at Northeast Ohio Medical University. The National Science Foundation Major Research Instrumentation Grant (CHE-0923398) supported the requisition of the ABSciex QTrap 5500 mass spectrometer instrument which was used in the optimization studies. The authors also thank Dr. Sameh Helmy for his contribution to finding a pertinent literature article to support the hypothesized chromatographic mechanism.
Funding Information:
The authors thank Dr. Tariq M. Haqqi for use of the LC-MS/MS instrumentation in his laboratory at Northeast Ohio Medical University, Rootstown, Ohio. This work was conducted with support from the Faculty and Research Development Program and the Dissertation Research Award program at Cleveland State University. This work was also supported by the Neurodegenerative Diseases and Aging Research Focus Area at Northeast Ohio Medical University. The National Science Foundation Major Research Instrumentation Grant (CHE-0923398) supported the requisition of the ABSciex QTrap 5500 mass spectrometer instrument which was used in the optimization studies. The authors also thank Dr. Sameh Helmy for his contribution to finding a pertinent literature article to support the hypothesized chromatographic mechanism.
Publisher Copyright:
© 2021 The Author(s). Published with license by Taylor & Francis Group, LLC.
PY - 2021
Y1 - 2021
N2 - A LC-MS/MS technique separated the bovine and mouse brain gangliosides monosialotetrahexosylgangliosides (GM1), disialotetrahexosylgangliosides (GD1a), trisialotetrahexosylgangliosides (GT1b) and tetrasialotetrahexosylgangliosides (GQ1b) using a phenyl-hexyl HPLC column and employing a linear methanol gradient in water, which is 0.028% in ammonium hydroxide. The gangliosides were separated according to sialic acid class, and within a particular class, gangliosides having different ceramide carbon chain lengths were also separated. All gangliosides of a particular sialic acid class eluted in characteristic retention time windows in the order of GQ1b, (earliest), GT1b, GD1a, and GM1 (latest). Within each specific retention time window for a particular ganglioside class, gangliosides were separated in the order of increasing ceramide carbon chain length. The phenyl-hexyl column separation of gangliosides is advantageous over established hydrophilic interaction and conventional reversed-phase chromatography techniques, in that the former separates gangliosides according to sialic acid class but not ceramide composition and the latter distributes all the sialic acid ganglioside classes throughout the entire chromatogram. The mechanism of separation of the ganglioside sialic acid classes is proposed to be a π-electron repulsion of negatively-charged gangliosides by the column phenyl moiety.
AB - A LC-MS/MS technique separated the bovine and mouse brain gangliosides monosialotetrahexosylgangliosides (GM1), disialotetrahexosylgangliosides (GD1a), trisialotetrahexosylgangliosides (GT1b) and tetrasialotetrahexosylgangliosides (GQ1b) using a phenyl-hexyl HPLC column and employing a linear methanol gradient in water, which is 0.028% in ammonium hydroxide. The gangliosides were separated according to sialic acid class, and within a particular class, gangliosides having different ceramide carbon chain lengths were also separated. All gangliosides of a particular sialic acid class eluted in characteristic retention time windows in the order of GQ1b, (earliest), GT1b, GD1a, and GM1 (latest). Within each specific retention time window for a particular ganglioside class, gangliosides were separated in the order of increasing ceramide carbon chain length. The phenyl-hexyl column separation of gangliosides is advantageous over established hydrophilic interaction and conventional reversed-phase chromatography techniques, in that the former separates gangliosides according to sialic acid class but not ceramide composition and the latter distributes all the sialic acid ganglioside classes throughout the entire chromatogram. The mechanism of separation of the ganglioside sialic acid classes is proposed to be a π-electron repulsion of negatively-charged gangliosides by the column phenyl moiety.
KW - Gangliosides
KW - HPLC-MS/MS
KW - LC-MS/MS
KW - mass spectrometry
KW - phenyl-hexyl column
UR - http://www.scopus.com/inward/record.url?scp=85103587017&partnerID=8YFLogxK
U2 - 10.1080/10826076.2020.1856136
DO - 10.1080/10826076.2020.1856136
M3 - Article
AN - SCOPUS:85103587017
SN - 1082-6076
VL - 44
SP - 114
EP - 125
JO - Journal of Liquid Chromatography and Related Technologies
JF - Journal of Liquid Chromatography and Related Technologies
IS - 1-2
ER -