Determination of serum high- and low-density lipoprotein cholesterol by ultracentrifugation-high performance liquid chromatography--improvement and verification of β-quantification method
10.3760/cma.j.issn.1009-8158.2019.08.008
- VernacularTitle:超速离心-高效液相色谱法测定高、低密度脂蛋白胆固醇的研究——β定量法的改良与验证
- Author:
Siming WANG
1
;
Jun DONG
;
Hongxia LI
;
Hongna MU
;
Ruiyue YANG
;
Weiyan ZHOU
;
Wenxiang CHEN
Author Information
1. 北京医院 国家老年医学中心
- Keywords:
Cholesterol;
HDL;
Cholesterol;
LDL;
Ultracentrifugation;
Reference standards
- From:
Chinese Journal of Laboratory Medicine
2019;42(8):623-628
- CountryChina
- Language:Chinese
-
Abstract:
Objective To propose and validate a reduced volume β-quantification method to measure serum high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C). Methods The reduced volume β-quantification method involved separation of LDL and HDL by ultracentrifugation and preparation of HDL by chemical precipitation. The sampling and reconstitution of the bottom fractions were performed gravimetrically and sample volume was thus decreased from 5 to 0.8 ml. High performance liquid chromatography was used to determine the cholesterol concentration of bottom fractions and HDL-C in the supernatant. Serum levels of LDL-C depended on a calculation of bottom fractions cholesterol minus HDL-C. Results The total CVs for HDL-C and LDL-C were 0.65％ -1.75％ and 0.63％ -1.11％. The results of the developed method were consistent with the current reference method and well within the allowable bias for Cholesterol Reference Method Laboratory Network surveys. Conclusion A new method for the measurement of HDL-C and LDL-C has been established. This method requires a small amount of serum and is easy to operate, exhibiting a desirable precision and accuracy. It is reliable and can be used as a candidate reference method for HDL-C and LDL-C.
