Determination of Cholesterol in Lipoprotein Fractions by Agarose Gel Electrophoresis
- VernacularTitle:琼脂糖凝胶电泳法测定血清中各种脂蛋白胆固醇
- Author:
Sheng-Kai YAN
1
;
Feng-Qing REN
;
Yao-Hong SONG
;
Qi-Sui LIN
Author Information
1. Biochemistry PUMC Hospital CAMS and PUMC
- From:
Acta Academiae Medicinae Sinicae
2001;23(1):93-96
- CountryChina
- Language:Chinese
-
Abstract:
Objective To evaluate a single step electrophoresis for quantitative determination of cholesterol of high-, low-, very-low-density lipoprotein(HDL, LDL, VLDL) and fast pre-beta lipoprotein [lipoprotein (a), Lp(a)]. Methods Quantification of lipoprotein cholesterol was performed by enzymatic staining of cholesterol in a new agarose gel electrophoresis method that allows the separation of LDL, VLDL, HDL, and Lp(a) by Helena REP system. The results of electrophoresis method were compared with those by traditional method like PTA-Mg2＋ precipitation method for HDL-C, PVS precipitation method for LDL-C, and Immunoturbidimetric assay(ITA) method for Lp(a).Results Within-runs CV were 5.16% ～ 7.46%,1.26% ～ 3.28% and 3.78% ～ 5.86% for VLDL-C, LDL-C and HDL-C, respectively. Between-runs CV were 8.35% ～ 11.25%, 2. 78% ～4. 08% and 4. 23% ～6. 36%, respectively. The linearity of this method was up to 10. 35 mmol/L total cholesterol. The recoveries were 90. 3%, 94. 3% and 89. 6%, respectively. No interference were observed when bilirubin( ＜ 342 μmol/L), hemoglobin( ＜ 20g/L) or triglyceride( ＜ 11.0 mmol/L) were added to pooled serum, respectively. There was good agreement between methods, with r = 0.9557 for HDL-C(electrophoresis method vs PTA-Mg2+ precipitation method),r = 0. 9609 for LDL-C (electrophoresis method vs PVS precipitation method) and r = 0. 9235 for Lp(a)-C (electrophoresis method) vs Lp(a) (ITA method). Conclusions The electrophoresis method offers a simple and inexpensive means of simultaneously measuring HDL-C, VLDL-C, Lp(a)-C and LDL-C.
