[通过琼脂糖凝胶电泳测定脂蛋白组分中的胆固醇]

[Determination of cholesterol in lipoprotein fractions by agarose gel electrophoresis].

作者信息

Yan S K, Ren F Q, Song Y H, Lin Q S

机构信息

Department of Clinical Biochemistry, PUMC Hospital, CAMS, PUMC, Beijing 100730, China.

出版信息

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2001 Feb;23(1):93-6.

PMID:12905830

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 mumol/L), hemoglobin(< 20 g/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.

摘要

目的

评估一种单步电泳法用于定量测定高、低、极低密度脂蛋白（HDL、LDL、VLDL）以及快速前β脂蛋白[脂蛋白(a)，Lp(a)]中的胆固醇。

方法

采用一种新的琼脂糖凝胶电泳法，通过对胆固醇进行酶染色来定量脂蛋白胆固醇，该方法可利用海伦娜REP系统分离LDL、VLDL、HDL和Lp(a)。将电泳法的结果与传统方法进行比较，如用于HDL-C的磷钨酸-镁（PTA-Mg2+）沉淀法、用于LDL-C的聚乙烯硫酸盐（PVS）沉淀法以及用于Lp(a)的免疫比浊法（ITA）。

结果

VLDL-C、LDL-C和HDL-C的批内变异系数分别为5.16%-7.46%、1.26%-3.28%和3.78%-5.86%。批间变异系数分别为8.35%-11.25%、2.78%-4.08%和4.23%-6.36%。该方法的线性范围高达总胆固醇10.35 mmol/L。回收率分别为90.3%、94.3%和89.6%。分别向混合血清中加入胆红素（<342 μmol/L）、血红蛋白（<20 g/L）或甘油三酯（<11.0 mmol/L）时，未观察到干扰。方法间具有良好的一致性，HDL-C（电泳法与PTA-Mg2+沉淀法）的r = 0.9557，LDL-C（电泳法与PVS沉淀法）的r = 0.9609，Lp(a)-C（电泳法）与Lp(a)（ITA法）的r = 0.9235。