Evolution of Methods for Measurement of HDL-Cholesterol: From Ultracentrifugation to Homogeneous Assays

Abstract

Background: Adoption of automated homogeneous assays for HDL-cholesterol (HDL-C) is increasing, driven by the need of clinical laboratories to cope with increasing workloads while containing costs. However, performance characteristics of homogeneous assays often differ in important aspects from those of the earlier precipitation methods. This review provides an overview of the new generation of homogeneous assays for HDL-C within the historical context of the evolution of methods and the efforts to standardize measurements of the lipoproteins. Approach: This is a narrative review based on method evaluations conducted in the laboratories of the authors as well as on relevant publications, especially comparative evaluation studies, from the literature. Publications considered here have been collected by the authors over the past 30 years of involvement as methods for HDL-C made the transition from their early use in lipid research laboratories to clinical laboratories and the recent emergence of homogeneous assays. Content: The presentation includes descriptions of methodologies, including homogeneous, precipitation, electrophoresis, and ultracentrifugation assays. Reference methods and recommended approaches for assessing accuracy are described. Accuracy and imprecision are summarized in the context of the National Cholesterol Education Program (NCEP) standards for analytical performance. The effects of interfering substances and preanalytical sources of variation are presented. Summary: Homogeneous assays have been shown to be reasonably well suited for use in routine clinical laboratories, generally meeting the NCEP criteria for precision, accuracy, and total error. However, discrepant results compared with the reference methods have been observed with some of the assays, and the sources of discrepancies are not well characterized. Some homogeneous reagents have not been thoroughly evaluated. At least three of the reagents have experienced successive adjustments in formulation; hence, the reagents may not yet be fully optimized. For these reasons, the homogeneous assays cannot be confidently recommended for use in long-term clinical trials and other research applications without thorough validation.

The role of lipase in blood lipoprotein metabolism and accumulation of lipids in oocytes of the Siberian sturgeon Acipenser baerii during maturation

“…In circulation, additional PL, cholesterol, and the minor apolipoproteins are transferred from TG-rich particles to these small, dense HDL particles, designated HDL 3 , converting them to larger and less dense spherical particles. Thus HDL 2 contain twice as many cholesterol molecules per unit of apolipoproteins compared with HDL 3 (Warnick et al, 2001). Therefore Increasing HL activity was promote large HDL 2 particles converting to HDL 3 , and would causing cholesterol carried by HDL decreased, which was just consistent with our result.…”

Apolipoprotein B/A-I and total cholesterol/high-density lipoprotein cholesterol ratios both predict cardiovascular events in the general population independently of nonlipid risk factors, albuminuria and C-reactive protein

“…HDL-C was measured using a homogeneous method (direct HDL, no. 7D67, AEROSET System; Abbott Laboratories, Abbott Park, IL, USA) [24]. Serum triglycerides were measured enzymatically.…”

Effects of rosuvastatin on lipids, lipoproteins and apolipoproteins in the dyslipidaemia of diabetes

“…than precipitation methods [30]. In the CARDS [14], HDL-C was measured by a precipitation method and the observed increase in HDL-C associated with atorvastatin 10 mg daily was a 1% increase compared with the placebo group.…”