Shotgun Sphingolipid Analysis of Human Aqueous Humor

Methods Mol Biol. 2018:1695:97-107. doi: 10.1007/978-1-4939-7407-8_10.

Abstract

This protocol provides a step-by-step guide to shotgun sphingolipid analysis of aqueous humor. We describe the Bligh and Dyer crude lipid extraction method and electrospray ionization tandem mass spectrometry (ESI-MS/MS) coupled with MZmine 2.21 data processing for identification and ratiometric quantitation of sphingosine, sphingosine-1-phosphate, sphingomyelin, and ceramide.

Keywords: Aqueous humor; Ceramide; Electrospray ionization-tandem mass spectrometry (ESI-MS/MS); MZmine 2.21; Shotgun lipidomics; Sphingolipids; Sphingomyelin; Sphingosine; Sphingosine-1-phosphate.

MeSH terms

  • Aqueous Humor / chemistry*
  • Ceramides / analysis
  • Humans
  • Lysophospholipids / analysis
  • Software
  • Spectrometry, Mass, Electrospray Ionization / methods
  • Sphingolipids / analysis*
  • Sphingomyelins / analysis
  • Sphingosine / analogs & derivatives
  • Sphingosine / analysis
  • Tandem Mass Spectrometry / methods

Substances

  • Ceramides
  • Lysophospholipids
  • Sphingolipids
  • Sphingomyelins
  • sphingosine 1-phosphate
  • Sphingosine