Analysis of aminophospholipid molecular species by methyl-beta-cyclodextrin modified micellar electrokinetic capillary chromatography with laser-induced fluorescence detection.
Micellar electrokinetic capillary chromatography (MEKC) with laser-induced fluorescence detection is used for the analysis of three classes of aminophospholipids: phosphatidylethanolamine (PE), phosphatidylserine (PS), and lysophosphatidylethanolamine (LPE) molecular species. 3-(2-Furoyl) quinoline-2-carboxaldehyde (FQ), a fluorogenic dye, was employed for labeling of these phospholipids. The FQ-labeled lipid species were then separated by sodium deoxycholate MEKC modified with methyl-beta-cyclodextrin. Baseline resolution of each class of phospholipids was achieved within 7 min. The migration time in each class increased with the carbon number of their side aliphatic chain. Separation efficiencies of approximately 3x10(5) plates were observed for most of these species. Concentration detection limits (3 sigma) were from 10(-9) to 10(-10) M for PE and LPE species and from 10(-8) to 10(-9) M for PS species. The relative standard deviations for migration time and peak area were less than 0.9% and 4.5%, respectively, for seven PE species. This method was applied to the separation of PE isolated from HT29 human colon cancer cells and roughly 30 PE species were resolved.
Department of Chemistry, University of Washington, Seattle, WA 98195, USA.
Le
L
Zhang
Shen
S
Hu
Lillian
L
Cook
Norman J
NJ
Dovichi
D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D003110:Colonic Neoplasms / Q000737:chemistry / Q000473:pathology; D003505:Cyclodextrins / Q000737:chemistry*; D003840:Deoxycholic Acid; D019075:Electrophoresis, Capillary / Q000379:methods; D005456:Fluorescent Dyes; D005470:Fluorometry; D006801:Humans; D007202:Indicators and Reagents; D007834:Lasers; D008246:Lysophospholipids / Q000032:analysis; D010714:Phosphatidylethanolamines / Q000032:analysis; D010718:Phosphatidylserines / Q000032:analysis; D010743:Phospholipids / Q000032:analysis*; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured; D047392:beta-Cyclodextrins
D003505:Cyclodextrins; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D008246:Lysophospholipids; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; D010743:Phospholipids; D047392:beta-Cyclodextrins; C008301:lysophosphatidylethanolamine; C108732:methyl-beta-cyclodextrin; D003840:Deoxycholic Acid