TY - JOUR
T1 - Direct quantitation of psychosine from alkaline-treated lipid extracts with a semi-synthetic internal standard
AU - Jiang, Xuntian
AU - Yang, Kui
AU - Han, Xianlin
PY - 2009
Y1 - 2009
N2 - Psychosine is an important bioactive sphingolipid metabolite and plays an essential role in the pathogenesis of Krabbe's disease. Herein, we extended shotgun lipidomics for the characterization and quantitation of psychosine in alkaline-treated crude lipid extracts by using neutral loss scan of 180 amu (i.e., galactose) in the positive-ion mode. Specifically, we semi-synthesized N,N-dimethyl psychosine and used it as an internal standard for quantitation of psychosine. After characterization of the fragmentation patterns of psychosine and the selected internal standard and optimization of the experimental conditions, we demonstrated that a broad linear dynamic range for the quantitation of psychosine and a limit of detection at a concentration of low fmol/ml were achieved using this approach. The developed method is generally simpler and more efficient than other previously reported methods. Multiple factors influencing quantitation of psychosine were extensively examined and/or discussed. The levels of psychosine in diabetic mouse nerve tissue samples were determined by the developed methodology. Collectively, the developed approach, as a new addition to the shotgun lipidomics technology, will be extremely useful for understanding the pathways/networks of sphingolipid metabolism and for exploring the important roles of psychosine in a variety of physiological and pathological conditions.
AB - Psychosine is an important bioactive sphingolipid metabolite and plays an essential role in the pathogenesis of Krabbe's disease. Herein, we extended shotgun lipidomics for the characterization and quantitation of psychosine in alkaline-treated crude lipid extracts by using neutral loss scan of 180 amu (i.e., galactose) in the positive-ion mode. Specifically, we semi-synthesized N,N-dimethyl psychosine and used it as an internal standard for quantitation of psychosine. After characterization of the fragmentation patterns of psychosine and the selected internal standard and optimization of the experimental conditions, we demonstrated that a broad linear dynamic range for the quantitation of psychosine and a limit of detection at a concentration of low fmol/ml were achieved using this approach. The developed method is generally simpler and more efficient than other previously reported methods. Multiple factors influencing quantitation of psychosine were extensively examined and/or discussed. The levels of psychosine in diabetic mouse nerve tissue samples were determined by the developed methodology. Collectively, the developed approach, as a new addition to the shotgun lipidomics technology, will be extremely useful for understanding the pathways/networks of sphingolipid metabolism and for exploring the important roles of psychosine in a variety of physiological and pathological conditions.
KW - Alzheimer's disease
KW - Diabetes mellitus
KW - Electrospray ionization mass spectrometry
KW - Krabbe's disease
KW - Shotgun lipidomics
KW - Sphingolipid metabolism
KW - Sphingolipidome
UR - http://www.scopus.com/inward/record.url?scp=63449110406&partnerID=8YFLogxK
U2 - 10.1194/jlr.D800036-JLR200
DO - 10.1194/jlr.D800036-JLR200
M3 - Article
C2 - 18753677
AN - SCOPUS:63449110406
SN - 0022-2275
VL - 50
SP - 162
EP - 172
JO - Journal of Lipid Research
JF - Journal of Lipid Research
IS - 1
ER -