Rapid high performance liquid chromatography-high resolution mass spectrometry methodology for multiple prenol lipids analysis in zebrafish embryos.
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.
Department of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy.
Chiara
C
Martano
Department of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy.
Vera
V
Mugoni
Department of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy.
Federica
F
Dal Bello
Department of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Laboratory of Endothelial Molecular Biology, Vesalius Research Center, VIB, Department of Oncology, University of Leuven, Leuven, Belgium.
Massimo M
MM
Santoro
Department of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy. Electronic address: claudio.medana@unito.it.
Claudio
C
Medana
D000818:Animals; D002784:Cholesterol / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004625:Embryo, Nonmammalian / Q000378:metabolism; D045782:Hemiterpenes; D013058:Mass Spectrometry / Q000379:methods; D009154:Mutation; D000439:Pentanols / Q000032:analysis*; D014451:Ubiquinone / Q000032:analysis; D014810:Vitamin E / Q000032:analysis; D010837:Vitamin K 1 / Q000032:analysis; D024482:Vitamin K 2 / Q000032:analysis; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism*; D024502:alpha-Tocopherol / Q000032:analysis
D045782:Hemiterpenes; D000439:Pentanols; D024482:Vitamin K 2; D014451:Ubiquinone; D014810:Vitamin E; C009034:prenol; D010837:Vitamin K 1; D002784:Cholesterol; D024502:alpha-Tocopherol
C4 reverse phase; HPLC–HRMS; Prenol lipids; Zebrafish