Quantification of lipoprotein (LP) particles was performed using ultracentrifugation (UC) as previously described (1). Seven different lipoprotein molecules including cholesterol (chol), triglycerides (tg), cholesterol ester (chole), free cholesterol (fchol), phospholipids (phosl), apolipoprotein A (apoA) and apolipoprotein B (apoB) were quantified in all or in some of the LP main fractions (VLDL, IDL, HDL, LDL) and/or in their subfractions (HDL2a, HDL2b, HDL3, LDL1, LDL2, LDL3, LDL4, LDL5, LDL6). Fractionation was done by sequential centrifugation of 3 mL EDTA plasma using an
Optima L-80 XP ultracentrifuge with a fixed angle rotor type 50.4
Ti (Beckman Coulter, Inc., US). The UC process was initiated immediately after the fasting plasma sample was collected, and it was completed over a period of 8 days. A detailed description of the separation steps can be found in (1). Immediately after the fractionation step, all subfractions were frozen and stored at -80⁰C for later analysis.
Colorimetric and turbidimetric assays were performed on an
ABX Pentra 400 analyzer (
ABX Pentra; Horiba ABX, Montpellier, France) to determine the plasma, main class and subclass concentrations of total chol, tg, apoA and apoB (
ABX Pentra; Horiba Medical, France). Free cholesterol and phoslp were determined using colorimetric and turbidimetric assays (MTI Diagnostics, Germany).
Khakimov B., Hoefsloot H.C.J., Mobaraki N., Aru V., Kristensen M., Lind M.V., Holm L., Castro‐Mejía J.L., Nielsen D.S., Jacobs D.M., Smilde A.K., & Engelsen S.B. (2021). Human blood lipoprotein predictions from 1H NMR spectra: protocol, model performances and cage of covariance.