TY - JOUR
T1 - Alkaline methanolysis of lipid extracts extends shotgun lipidomics analyses to the low-abundance regime of cellular sphingolipids
AU - Jiang, Xuntian
AU - Cheng, Hua
AU - Yang, Kui
AU - Gross, Richard W.
AU - Han, Xianlin
N1 - Funding Information:
This work was supported by NIA Grant R01 AG23168, NIA Grant R01 AG31675, NIH Grant P01 HL57278, and the Neurosciences Education and Research Foundation.
PY - 2007/12/15
Y1 - 2007/12/15
N2 - Sphingolipids that contain a sphingoid base are composed of hundreds to thousands of distinct compounds, many of which serve as lipid regulators of biological functions. The global analysis of the large number of low-abundance sphingolipid molecular species has been hampered in many cases by the sphingolipid molecular species being overwhelmed by the quantity of other classes of lipid (e.g., glycerophospholipid) molecular species present, thereby imposing severe restrictions on the dynamic range of their measurement using shotgun lipidomics. Herein, we developed a facile approach in which the sphingolipids of cellular extracts were dramatically enriched by direct alkaline methanolysis of lipid extracts followed by extraction to remove the large majority of other endogenous lipid classes. Through direct infusion of the resultant enriched solution, we identified and quantitated a variety of very-low-abundance sphingolipid classes (e.g., sphingosine, psychosine, and lysosphingomyelin) and molecular species (e.g., sphingomyelin) using electrospray ionization mass spectrometry (i.e., shotgun sphingolipidomics). Accordingly, through utilization of these facile enrichment techniques, direct penetrance into the sphingolipidomes has been greatly extended, facilitating new insights into their metabolism and signaling functions in biological systems.
AB - Sphingolipids that contain a sphingoid base are composed of hundreds to thousands of distinct compounds, many of which serve as lipid regulators of biological functions. The global analysis of the large number of low-abundance sphingolipid molecular species has been hampered in many cases by the sphingolipid molecular species being overwhelmed by the quantity of other classes of lipid (e.g., glycerophospholipid) molecular species present, thereby imposing severe restrictions on the dynamic range of their measurement using shotgun lipidomics. Herein, we developed a facile approach in which the sphingolipids of cellular extracts were dramatically enriched by direct alkaline methanolysis of lipid extracts followed by extraction to remove the large majority of other endogenous lipid classes. Through direct infusion of the resultant enriched solution, we identified and quantitated a variety of very-low-abundance sphingolipid classes (e.g., sphingosine, psychosine, and lysosphingomyelin) and molecular species (e.g., sphingomyelin) using electrospray ionization mass spectrometry (i.e., shotgun sphingolipidomics). Accordingly, through utilization of these facile enrichment techniques, direct penetrance into the sphingolipidomes has been greatly extended, facilitating new insights into their metabolism and signaling functions in biological systems.
KW - Electrospray ionization mass spectrometry
KW - Lysosphingomyelin
KW - Psychosine
KW - Shotgun lipidomics
KW - Shotgun sphingolipidomics
KW - Sphingolipidome
KW - Sphingomyelin
KW - Sphingosine
UR - http://www.scopus.com/inward/record.url?scp=35548980403&partnerID=8YFLogxK
U2 - 10.1016/j.ab.2007.08.019
DO - 10.1016/j.ab.2007.08.019
M3 - Article
C2 - 17920553
AN - SCOPUS:35548980403
SN - 0003-2697
VL - 371
SP - 135
EP - 145
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -