Prince:BYU Lipidomics Team

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Basic Protocol

• Get sample from almost any source
• Extraction (both methods use Folch reagent [2:1 chloroform:methanol])
  • use Kaluzne method
  • trying to implement Bligh & Dyer (gold standard)
  • MTBE Extraction [1]
• Mass Spectrometry
  • ESI-Agilent TOF
  • Tandem MS-Q-Star (8-10am)
• Analysis
  • Matlab (particular toolbox?) PCA analysis

Main analytes are phosphatidylcholine and phosphatidylethanolamine. Analytes in (-) mode would be phosphatidylserines and phosphatidylethanolamines (which are expected to have stronger signal in - mode). Typical number of analytes?

FAME-GC associated with Craig Thulin and UVU.

Literature

Great review on the basics of ESI and quantitative, shotgun lipidomics. [2]

Practical aspects of ESI. [3]

Review covering many aspects of lipidomics. [4]

Group Members

Current

• Mary Blackburn
• Mackay Merrill
• Dean Flanders
• Landon Weist
• Rob Roden
• Greg Jackson
• Daniel Edmondson
• Matthew Linford
• Craig Thulin
• Steven Wood
• John Prince

Former or Associated

• Jennifer MacDonald
• Jonathan Lee
• Vinod Chaudhary
• Steven Herron

Relevant Publications

1. Dang L, White DW, Gross S, Bennett BD, Bittinger MA, Driggers EM, Fantin VR, Jang HG, Jin S, Keenan MC, Marks KM, Prins RM, Ward PS, Yen KE, Liau LM, Rabinowitz JD, Cantley LC, Thompson CB, Vander Heiden MG, and Su SM. Cancer-associated IDH1 mutations produce 2-hydroxyglutarate. Nature. 2009 Dec 10;462(7274):739-44. DOI:10.1038/nature08617 |

   This has a great protocol for negative ion mode extraction and analysis on the Orbitrap.