Capillary Electrophoresis of Phospholipids with Indirect Photometric Detection

• Published in: Journal of chromatographic science Vol. 36; no. 8; pp. 395 - 400
• Main Authors: Haddadian, Fereshteh, Shamsi, Shahab A., Schaeper, James P., Danielson, Neil D.
• Format: Journal Article
• Language: English
• Published: Niles, IL Oxford University Press 01.08.1998; Preston Publications
• Subjects: 1-Propanol; Acetonitriles; Adenosine Monophosphate; Analytical chemistry; Analytical, structural and metabolic biochemistry; Animals; Anions; Biological and medical sciences; Brain Chemistry; Chemistry; Chromatographic methods and physical methods associated with chromatography; Electrophoresis, Capillary - methods; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; Glycerolipids, phospholipids; Glycine max - chemistry; Lipids; Methanol; Other biological molecules; Other chromatographic methods; Phospholipids - isolation & purification; Solvents; Surface-Active Agents; Swine; Zone electrophoresis; Phase composition; Chromatographic retention; Capillary electrophoresis; Non ionic surfactant; Phospholipid; Experimental study; Complex lipid; Mobile phase; Organic solvent; Ultraviolet detector; Indirect method; Detection limit
• ISSN: 0021-9665; 1945-239X
• DOI: 10.1093/chromsci/36.8.395

The capillary electrophoresis (CE) separation of different anionic phospholipid classes including phosphatidic acids (PA), phosphatidylserine, phosphatidylinositol, phosphatidylglycerol (PG), and cardiolipin using indirect detection with adenosine monophosphate (AMP) is described. A standard mixture of PAs (C14, C16, and C18) can be separated in 10 min by CE using 5mM AMP and 100mM boric acid in 10% water—80% methanol—10% acetonitrile. Although nonionic surfactants such as Brij 35 can improve the CE resolution of PAs, the separation time and the baseline noise are both increased. Optimization of the organic solvent in the running electrolyte is important. Methanol provides faster electroosmotic flow than propanol, and 10% acetonitrile effectively reduces migration time further by a factor of 1.4–2.2, depending on the phospholipid. The concentration limit of detection ranges from approximately 2 to 6 mg/L, and the mass limit of detection is as low as 21 pg. Linearity from 19 to 100 mg/L is established for cardiolipin and C16-PG. Phospholipids in soybean and brain extract samples could be profiled.