YAKHAK HOEJI (약학회지)

The Pharmaceutical Society of Korea (대한약학회)
권호 표지

Determination of Monoglycoceramides in Biological Samples using Enzymatic Deacylation and Reverse-phase HPLC

역상HPLC컬럼을 이용한 생체 내 단당세라마이드 분석

  • Choi, Mi-Hwa (College of Pharmacy and CBITRC, Chungbuk National University) ;
  • Choi, Kyeong-Mi (College of Pharmacy and CBITRC, Chungbuk National University) ;
  • Ji, So-Young (College of Pharmacy and CBITRC, Chungbuk National University) ;
  • Lee, Youn-Sun (College of Pharmacy and CBITRC, Chungbuk National University) ;
  • Cho, Ju-Hyun (College of Pharmacy and CBITRC, Chungbuk National University) ;
  • Lee, Yong-Moon (College of Pharmacy and CBITRC, Chungbuk National University) ;
  • Yun, Yeo-Pyo (College of Pharmacy and CBITRC, Chungbuk National University) ;
  • Yoo, Hwan-Soo (College of Pharmacy and CBITRC, Chungbuk National University)
  • Received : 2010.06.01
  • Accepted : 2010.09.02
  • Published : 2010.10.31
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Abstract

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 $141.5{\pm}5$ pmol/$1{\times}10^6$ cells and $3.9{\pm}0.3{\mu}M$, 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.

Keywords

References

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