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琼脂糖凝胶电泳法测定血清中各种脂蛋白胆固醇 被引量:3

Determination of Cholesterol in Lipoprotein Fractions by Agarose Gel Electrophoresis
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摘要 对琼脂糖凝胶电泳法同时测定血清极低密度脂蛋白( VLDL)、低密度脂蛋白( LDL)、高密度脂蛋白( HDL)和脂蛋白 (a)[Lp(a)]的胆固醇 [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),然后结合胆固醇酶染色法同时测定 VLDL- C、 LDL- C、 HDL- C和 Lp(a)- C的含量。分析了该法的精密度、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)的免疫透射比浊法( ITA法)进行比较。结果电泳法测定 VLDL- C、 LDL- C和 HDL- C的批内 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 1.04~ 10.35 mmol/L,基本不受溶血、黄疸和脂血干扰。 VLDL- C、 LDL- C和 HDL- C的平均回收率分别为 90.3%、 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 Lp(a)的 r为 0.9235。以该法检测北京地区 98例健康人, HDL- C、 VLDL- C、 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(< 342μ mol/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.
出处 《中国医学科学院学报》 CAS CSCD 北大核心 2001年第1期93-96,共4页 Acta Academiae Medicinae Sinicae
关键词 脂蛋白 胆固醇 琼脂糖凝胶电泳法 测定 lipoprotein cholesterol electrophoresis
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