出版社：American Society for Biochemistry and Molecular Biology
摘要：Here, we have extended shotgun lipidomics for the characterizationand quantitation of sphingosine-1-phosphate (S1P) and dihydrosphingosine-1-phosphate(DHS1P) in crude lipid extracts in the presence of ammoniumhydroxide by using precursor ion scanning of m/z 79.0 (correspondingto [PO3]–) in the negative-ion mode. It is demonstratedthat a broad linear dynamic range for the quantitation of bothS1P and DHS1P and a detection limit at low amol/µl concentrationare achieved using this approach. The developed method for thequantitation of sphingoid base-1-phosphates is generally simplerand more efficient than other previously published methods.Multiple factors influencing the quantitation of sphingoid base-1-phosphates,including ion suppression, extraction efficiency, and potentialoverlapping with other molecular species, were examined extensivelyand/or are discussed. Mass levels of S1P and DHS1P in multiplebiological samples, including human plasma, mouse plasma, andmouse brain tissues (e.g., cortex, cerebellum, spinal cord,and brain stem), were determined by the developed methodology.Accordingly, this technique, as a new addition to shotgun lipidomicstechnology, will be extremely useful for understanding the pathwaysof sphingolipid metabolism and for exploring the important rolesof sphingoid base-1-phosphates in a wide range of physiologicaland pathological studies.Supplementary key words dihydrosphingosine-1-phosphate • electrospray ionization mass spectrometry • brain tissues • sphingolipid metabolism
