In:
Evidence-Based Complementary and Alternative Medicine, Hindawi Limited, Vol. 2012 ( 2012), p. 1-9
Abstract:
Sphingomyelin is the most abundant sphingolipid in mammalian cells and is mostly present in the plasma membrane. A new analytical method using high-performance liquid chromatography (HPLC) was developed to quantify sphingomyelin in mouse plasma and tissues, 3T3-L1 cells, rat aortic smooth muscle cells, and HT-29 cells. Sphingomyelin and dihydrosphingomyelin, an internal standard, were separated by high-performance thin-layer chromatography and simultaneously hydrolyzed with sphingolipid ceramide N -deacylase and sphingomyelinase to release sphingosine and dihydrosphingosine, respectively. Sphingomyelin content was measured by HPLC following o -phthalaldehyde derivatization. Sphingomyelin concentrations in 3T3-L1 cells, rat aortic smooth muscle cells, and HT-29 cells were 60.10 ± 0.24 , 62.69 ± 0.08 , and 58.38 ± 0.37 pmol/ μ g protein, respectively, whereas those in brain, kidney, and liver of ICR mice were 55.60 ± 0.43 , 43.75 ± 0.21 , and 22.26 ± 0.14 pmol/ μ g protein. The sphingomyelin concentration in mouse plasma was 407.40 ± 0.31 μ M. The limits of detection and quantification for sphingomyelin were 5 and 20 pmol, respectively, in the HPLC analysis with fluorescence detection. This sensitivity was sufficient for analyzing sphingomyelin in biological samples. In conclusion, this analytical method is a sensitive and specific technique for quantifying sphingomyelin and was successfully applied to diverse biological samples with excellent reproducibility.
Type of Medium:
Online Resource
ISSN:
1741-427X
,
1741-4288
Language:
English
Publisher:
Hindawi Limited
Publication Date:
2012
detail.hit.zdb_id:
2148302-4
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