琼脂糖凝胶电泳法测定血清中各种脂蛋白胆固醇

目的：对琼脂糖凝胶电脉法同时测定血清极密度脂蛋白（VLDL），低密度脂蛋白（LDL），高密度脂蛋白（HDL）和脂蛋白（a)Lp(a）的胆固 即VLDL－C，LDL－C，HDL－C和Lp(a)-C含量的方法进行评价。方法：采用Helena REP全自动快速电泳系统分离血清VLDL，LDL，HDL和Lp(a)，然后结合胆固醇染色法同时测定VLDL－C，LDL－C，HDL－C和Lp(a)-C的含量，分别了该法的精密度，准确度和干扰因素，并将该法与传统方法如测定HDL－C的磷钨酸－镁沉淀法（PTA－Mg2 )法，测定LDL－C的聚乙烯硫酸沉淀法（PVS法）及测定Lp(a)的免疫秀射比浊法（ITA法）进行比较，结果：电泳法测定VLDL－C，LDL－C和HLD－C批内CV分别为5．16％－7．46％,1.26％－3．28%，3．78－5．86％；批间CV分别为8．35％－11．25％，2．78％－4．08％，4．23％－6．36％。检测线性范围总胆固醇为94．3％和89．6。电泳法与传统法测定LDL－C，HDL－C的相关系数（r)分别为0．9609，0．9557，电泳法测定Lp(a)-C与ITA法测定Lq(a)的r为9235，以该法检测北京地区98例健康人，HDL－C，VLDL－C，LDL－C，Lp(a)－C参考范围分别为0．95－2．10mmol/L,0.07-0.46mmol/L,1.77-2.87mmol/L,0-0.22mmol/L。结论：电泳法测定值与传统法一致，电泳法操作简便，快速经济，可同时测定4种脂蛋白胆固醇，适合于临床常规及研究使用。

Determination of cholesterol in lipoprotein fractions by agarose gel electrophoresis

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.