HDL-Particles Separation Employing Different Precipitating Agents: Functional Properties of the Impact of Chemical Precipitation on Lipoprotein Particle-Size and Paraoxonase-1-Activity

Abstract

First-generation methods used to quantify HDL-cholesterol have been used for decades. Therefore, first-generation precipitatingreagents still in use, the chemical impact on HDL-particles functionality (structure/activity-relationship) is not well-documented. The study aim was to identify the most efficient precipitating-reagent for lipoprotein separation to be used in the HDL functionalevaluation. A cross-sectional design with convenience sampling was conducted. Plasma samples from 40 individuals, both genders, were evaluated. Biochemical analysis was performed after using different chemical precipitants (e.g. polyethylene glycol-PEG (P1), phosphotungstic-acid (P2), and dextran-sulfate/MgCl 2 (P3)) to evaluate HDL-particle-size and PON1-activity. Parametric and non-parametric tests were performed using GraphPad-Prism 5.01 (USA). Differences were considered significant when a p-value < 0.05 were obtained. The PON1-activity comparison differed after NaCl stimulation (p < 0.001, paired t-test) and among chemical precipitant treatments. The PON1-activity differed after sample precipitation with P1 when compared to P2, and P3 (p < 0.05; ANOVA), before and after stratification for dyslipidemia group. The HDL particle-size comparison did not differ, but on the other hand, polydispersity differed after sample precipitation with P3 when compared to P1 or P2 (p < 0.05; ANOVA). Dextran-Sulfate/MgCl 2 was a better chemical precipitant for PON1-activity, and PEG for HDL-particle-size, respectively. The HDL functional evaluation depends on the appropriately selected chemical precipitant.

Unlocking the Potential of Stingray (Dasyatis sephen) Extract: A Novel Investigation into EPA and DHA Effects on Lipid Metabolism in High-Fat Diet-Fed Wistar Rats

“…We obtained a clean supernatant by centrifuging the mixture at 4000 rpm for 20 minutes. The absorbance of this supernatant, signifying the HDL-cholesterol level, was determined using spectrophotometry at a wavelength of 546 nm (16,17).…”