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KMID : 0370220100540050354
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Yakhak Hoeji
2010 Volume.54 No. 5 p.354 ~ p.361
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Determination of Monoglycoceramides in Biological Samples using Enzymatic Deacylation and Reverse-phase HPLC
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Choi Mi-Hwa
Choi Kyeong-Mi
Ji So-Young
Lee Youn-Sun
Cho Ju-Hyun
Lee Yong-Moon
Yun Yeo-Pyo
Yoo Hwan-Soo
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Abstract
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Glycosphingolipids are structural components of mammalian cell membranes and are involved in essential cellular physiology such as cell-cell interaction, recognition, transmembrane signaling, proliferation and cell death. In this study, the simple quantitative method of monoglycoceramides-containing glucosylceramide and galactosylceramide was developed. The glycosylceramides extracted from culture cells and rat plasma were resolved by TLC, deacylated by SCDase and analyzed by HPLC-fluorescence detector at an excitation wavelength of 340 nm and an emission wavelength of 455 nm. Limit of detection was approximately 0.1 pmol and limit of quantification was about 1 pmol for both monoglycoceramide standards. The recoveries of standard glucosylceramides from intra- and inter-day assays were 113.8 and 88.8% and those of galactosylceramides were 110.7 and 123.9%, respectively. The monoglycoceramide contents of SW-620 cells and rat plasma were pmol/ cells and , respectively. The present analytical method provides a reproducible quantification and total content of monoglycoceramide which may be as a potential biomarker for lipid imbalance-related human diseases.
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KEYWORD
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monoglycoceramide, glucosylceramide, galactosylceramide, SCDase, HPLC, SW-620 cells
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