ZFIN ID: ZDB-PUB-150819-4
Rapid high performance liquid chromatography-high resolution mass spectrometry methodology for multiple prenol lipids analysis in zebrafish embryos
Martano, C., Mugoni, V., Dal Bello, F., Santoro, M.M., Medana, C.
Date: 2015
Source: Journal of chromatography. A   1412: 59-66 (Journal)
Registered Authors: Mugoni, Vera, Santoro, Massimo
Keywords: C4 reverse phase, HPLC–HRMS, Prenol lipids, Zebrafish
MeSH Terms:
  • Animals
  • Cholesterol/analysis
  • Chromatography, High Pressure Liquid/methods
  • Embryo, Nonmammalian/metabolism
  • Mass Spectrometry/methods
  • Mutation
  • Pentanols/analysis*
  • Ubiquinone/analysis
  • Vitamin E/analysis
  • Vitamin K 1/analysis
  • Vitamin K 2/analysis
  • Zebrafish/genetics
  • Zebrafish/metabolism*
  • alpha-Tocopherol/analysis
PubMed: 26283533 Full text @ J. Chromatogr. A
The analysis of lipid molecules in living organism is an important step in deciphering metabolic pathways. Recently, the zebrafish has been adopted as a valuable animal model system to perform in vivo metabolomics studies, however limited methodologies and protocols are currently available to investigate zebrafish lipidome and even fewer to analyze specific classes of lipids. Here we present an HPLC-HRMS based method to rapidly measure multiple prenol lipid molecules from zebrafish tissues. In particular, we have optimized our method for concurrent detection of ubiquinones (Coenzyme Q6, Coenzyme Q9, Coenzyme Q10), cholesterol, vitamin E (α-tocopherol), vitamin K1 and vitamin K2. The purpose of this study was to compare different ionization modes, mobile phases and stationary phases in order to optimize lipid molecules separation. After HPLC-HRMS parameters selection, several extraction conditions from zebrafish embryos were evaluated. We assessed our methodology by quantitation of analytical recovery on zebrafish extracts from wild-type or zebrafish mutants (barolo) affected by impaired biosynthesis of ubiquinones.