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A more recent version of this article appeared on March 1, 2006

Papers In Press, published online ahead of print December 21, 2005
J. Lipid Res., doi:10.1194/jlr.D500040-JLR200
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Submitted on November 11, 2005
Revised on December 5, 2005
Accepted on December 7, 2005

Rapid, specific and sensitive measurements of plasma sphingomyelin and phosphatidylcholine

Mohammad Reza Hojjati and Xian-Cheng Jiang

Anatomy and Cell Biology, SUNY, Brooklyn, NY 11203

Corresponding Author: xjiang{at}downstate.edu

Sphingomyelin (SM) and phosphatidylcholine (PC) are two major phospholipids on plasma lipoproteins. Their concentration is classically measured by lipid extraction, thin layer chromatograph, and phosphate determination on separated SM or PC spots. Here, we describe two rapid, specific and sensitive enzymatic measurements for both phospholipids. Plasma was incubated with bacterial sphingomyelinase (for SM measurement) or bacterial PC-specific phospholipase D (for PC measurement), alkaline phosphatase, choline oxidase, peroxidase, N-Ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline, and 4-aminoantipyrine for 45 minutes. A blue dye, with an optimal absorption at 595 nm, was generated. PC levels did not influence SM measurement and vice versa. The linear range for the SM measurement was 0.5 to 5 g and for PC it was 2.5 to 20 g. The mean percent recovery of SM was 98.0+5.2% and PC was 96.6+3.8%. The inter-assay coefficient of variation of the assay was 1.7+0.05% for SM and 3.1+0.13% for PC. These two new methods are amenable to automation and can be adopted for large-scale, high-throughput assays.


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