Reiko Kiyonami, David A. Peake and Andreas Huhmer
ASMS 2017 Poster
Purpose: Develop a robust and reproducible HPLC MS-MS method which enables untargeted lipid identification and targeted screening of thousands major lipid molecule for getting estimated concentrations of identified lipid species in a single HPLC MS-MS run on a Thermo Scientific™ Q Exactive™ HF-X hybrid quadrupole-Orbitrap™ mass spectrometer.
Methods: Analysis of lipid extracts from bovine heart and human plasma. An isotopically labeled lipid standard mixture (SPLASH™ Lipidomix® Mass Spec Standard, Avanti Polar Lipids) was spiked into each sample prior to lipid extraction. A large inclusion list (positive and negative ions) covering 4074 lipid molecular species from 14 major lipid classes was generated in-silico using the lipid database included in the Thermo Scientific™ LipidSearch™ software and used to direct the LC MS-MS data acquisition. LipidSearch pre-release software was used for lipid identification and quantitation. The estimated concentrations of identified lipid species from 14 major lipid classes were calculated relative to the known concentration of isotopic labeled standards included in the SPLASH mixture.
Results: Deep human plasma lipidome coverage was observed using the directed HPLC MS-MS workflow. More than 1000 lipid species were quantified with estimated concentrations using the isotopic labeled lipid standards from the Splash mixture.
Thermo Fisher Scientific