Related Experiment Video
Updated: May 7, 2026
High-Density Lipoprotein-Specific Phospholipid Efflux Assay
Published on: September 30, 2025
Development of a homogeneous assay for measurement of high-density lipoprotein-subclass cholesterol
Yasuki Ito1, Noriyuki Satoh, Takayoshi Ishii
1Reagent R&D department, Denka Seiken Co., Ltd., 1359-1, Kagamida, Kigoshi, Gosen-shi, Niigata 959-1695, Japan.
Background:
Several studies have suggested that measurement of high-density lipoprotein (HDL) 2 and HDL3 subfractions might be more useful for evaluating coronary risk than total HDL-cholesterol (C). However, methods of measuring HDL2 and HDL3 are quite laborious for general clinical use. Development of a quick and easy method of measuring HDL subfractions has been long-awaited.
Methods:
Triglyceride (TG) rich lipoproteins (TRLs), low-density lipoprotein (LDL), HDL2, and HDL3 were used for screening of surfactants and enzymes to react selectively with HDL3-C and to decompose other lipoproteins.
Results:
In order to develop HDL3-C homogeneous assay, polyoxyethylene styrenated phenyl ether derivative, for which the hydrophilic lipophilic balance (HLB) value is 13.6, was adopted as the most effective and specific surfactant for selection of HDL3 from HDL. Sphingomyelinase (SMase) reacted with TRLs and LDL preferentially, and decomposed them. HDL2-C was estimated by subtracting measured HDL3-C from total HDL-C, directly measured by homogeneous method. The homogeneous assay exhibited excellent correlations with the results of HDL3-C and HDL2-C measured by standard ultracentrifugation (R(2)=0.848 and 0.982, respectively).
Conclusions:
We established a rapid and effective, fully-automated assay for the measurement of HDL3-C. Furthermore, the subtraction of HDL3-C from total HDL-C allows concurrent determination of HDL2-C.
