Quantitative Analysis of Sphingomyelin by High-Performance Liquid Chromatography after Enzymatic Hydrolysis
Figure 1
Sphingomyelin quantification procedure and the HPLC chromatogram. (a) Sphingomyelin was simultaneously hydrolyzed with sphingomyelinase (SMase) and sphingolipid ceramide N-deacylase (SCDase), and the released sphingosine was derivatized with OPA followed by HPLC analysis. (b) Sphingomyelin was separated from lipid extracts of ICR mouse kidney tissue by high-performance thin-layer chromatography (HPTLC), simultaneously hydrolyzed with SMase and SCDase, and analyzed by HPLC following OPA derivatization. The peaks representing sphingomyelin and dihydrosphingomyelin (internal standard) occurred at 14.4 and 19.8 min, respectively.
