Plasma Lipid and Cholesterol Analysis

Plasma lipids were processed for measurement of total cholesterol, esterified cholesterol, HDL cholesterol, LDL-cholesterol, triglycerides and free fatty acids by enzymatic procedures as previously described (27 (link)). To obtain FPLC plasma cholesterol profiles, plasma (200 µl pooled from 2 mice) was fractionated using a Superose 6 column (Amersham Biosciences, GE Life Sciences, Pittsburgh, PA) on a Biologic DuoFlow FPLC system (Bio-Rad, Hercules, CA) as previously described (20 (link)). One-half ml fractions were collected and analyzed for total cholesterol content using enzymatic cholesterol reagents (Infinity Cholesterol Reagent TR13421, Thermo Scientific). Total plasma and fractionated plasma PON1 paraoxonase activity were measured using 1 mM Paraoxon (Supelco Analytical, Sigma Aldrich, St. Louis, MO) in 50 mM Tris/HCl pH 8.0, 1mM CaCl₂ as previously described (18 (link)).