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J. Biochem, 1984, Vol. 95, No. 6 1671-1675
© 1984 Japanese Biochemical Society

research-article

Analysis of Phospho- and Phosphonosphingolipids by High-Performance Liquid Chromatography

Osamu ITASAKA, Taro HORT, Kimiko SASAHARA, Yaeko WAKABAYASHI, Fumiko TAKAHASHI and Hae-ik RHEE1

Department of Chemistry, Faculty of Liberal Arts and Education Shiga University Otsu, Shiga 520

A simple and efficient method for the separation of phosphosphingolipids including phosphonosphingolipids by high-performance liquid chromatography is described. A mixture of authentic lipids consisting of sphingomyelin, ceramide phosphorylethanolamine, ceramide 2-aminoethylphosphonate, and ceramide N-methylaminoethylphosphonate was completely separated using a silica gel (Zorbax SIL) column with acetonitrile-methanol-water 72 : 40 : 10 (v/v) as eluting solvent. The elution of these sphingolipids was monitored directly with an ultraviolet spectromonitor at 207 nm. The practical limit of detection of each sphingolipid was about 0.2 µg or 0.3 nmol. Using this method, we found that from one to four different phosphonoand/or phosphosphingolipids in fresh-water shellfish can be routinely identified and reproducibly quantified.

1Present address: Kangwon National University, Chuncheon, Korea.


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