Micellar electrokinetic chromatography profiles of human high‐density lipoprotein phospholipids

• DOI: 10.1002/elps.201000474
• Publication Date: 2011-04-18T07:38:03Z
• Authors: Chin‐Pong Chong; Ting‐Yu Lin; Chia‐Liang Chang; Ying‐Ling Yang; Ming‐Hua Tsai; Yu‐Shan Yu; Mine‐Yine Liu
• Keywords: Micellar electrokinetic chromatography

Abstract A simple and fast micellar electrokinetic chromatography (MEKC) method was developed to investigate phospholipids isolated from human high‐density lipoproteins (HDL). To optimize the MEKC conditions, several factors including bile salt concentration organic modifier in separation buffer as well temperature have been examined. The optimal chosen a mixture of 50 mM salts, 30% v/v 1‐propanol 10 sodium phosphate (pH 8.5). applied voltage selected were 25 kV 40°C, respectively. Meanwhile, high‐salt stacking has performed for sample pre‐concentration enhance peak sensitivity. Several matrix injection time optimized. 100 NaCl 20% 1‐propanol, 32 s under pressure 0.5 psi. phospholipid standards lysophosphatidyl choline, phosphatidyl choline (PC), sphingomyelin, ethanolamine, inositol, serine phosphatidic acid studied using method. profile mixed showed good reproducibility. linear ranges PC sphingomyelin 0.025–1.2 0.025–2.0 mg/mL, limits detection 0.0156 0.0199 Using an internal standard, precision accuracy measured sphingomyelin. intraday interday quantitative analysis results. new used characterize native, vitro oxidized glycated HDL within 16 min. At absorbance 200 nm, two similar peaks observed native phospholipids, but three phospholipids. Interestingly, at 234 distinctively different profiles