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gc/ms for sphingosine

Ed Siefker via methods%40net.bio.net (by ebs15242 from gmail.com)
Tue Nov 21 14:42:49 EST 2017


I am interested in assaying sphingosine from cultured cells.  I have found
many methods for LC-MS.  Unfortunately, my lab only has a GC-MS.
I have been unable to find any discussion of GC-MS methods for sphingosine.

ELISA methods exist for sphingosine-1-phosphate, and sphingosine can be
phosphorylated easily enough.  I could subtract endogenous S1P from total
S1P, but endogenous S1P is present at much higher concentrations than
sphingosine, so I'm afraid my signal would be lost in the noise.

Phosphorylation with 32P followed by TLC and scintillation counting is an
option, but a last resort.

Are there approaches I haven't considered here, or papers where these approaches
have been successfully used?



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