用新型简易高效液相色谱法测定尿白蛋白
Determination of Urine Albumin by New Simple High-Performance Liquid Chromatography Method.
作者信息
Klapkova Eva, Fortova Magdalena, Prusa Richard, Moravcova Libuse, Kotaska Karel
机构信息
Department of Medical Chemistry and Clinical Biochemistry, 2nd Faculty of Medicine and University Hospital Motol, Charles University, Prague, Czech Republic.
出版信息
J Clin Lab Anal. 2016 Nov;30(6):1226-1231. doi: 10.1002/jcla.22007. Epub 2016 May 31.
BACKGROUND
A simple high-performance liquid chromatography (HPLC) method was developed for the determination of albumin in patients' urine samples without coeluting proteins and was compared with the immunoturbidimetric determination of albumin. Urine albumin is important biomarker in diabetic patients, but part of it is immuno-nonreactive.
METHODS
Albumin was determined by high-performance liquid chromatography (HPLC), UV detection at 280 nm, Zorbax 300SB-C3 column. Immunoturbidimetric analysis was performed using commercial kit on automatic biochemistry analyzer COBAS INTEGRA 400, Roche Diagnostics GmbH, Manheim, Germany.
RESULTS
The HLPC method was fully validated. No significant interference with other proteins (transferrin, α-1-acid glycoprotein, α-1-antichymotrypsin, antitrypsin, hemopexin) was found. The results from 301 urine samples were compared with immunochemical determination. We found a statistically significant difference between these methods (P = 0.0001, Mann-Whitney test).
CONCLUSION
New simple HPLC method was developed for the determination of urine albumin without coeluting proteins. Our data indicate that the HPLC method is highly specific and more sensitive than immunoturbidimetry.
背景
开发了一种简单的高效液相色谱（HPLC）方法，用于测定患者尿液样本中的白蛋白，且不存在共洗脱蛋白的情况，并与免疫比浊法测定白蛋白进行了比较。尿白蛋白是糖尿病患者重要的生物标志物，但其中一部分具有免疫非反应性。
方法
采用高效液相色谱（HPLC）、280nm紫外检测、Zorbax 300SB-C3柱测定白蛋白。使用商业试剂盒在德国曼海姆罗氏诊断有限公司的自动生化分析仪COBAS INTEGRA 400上进行免疫比浊分析。
结果
HLPC方法得到了充分验证。未发现与其他蛋白质（转铁蛋白、α-1-酸性糖蛋白、α-1-抗糜蛋白酶、抗胰蛋白酶、血红素结合蛋白）有明显干扰。将301份尿液样本的结果与免疫化学测定结果进行了比较。我们发现这些方法之间存在统计学显著差异（P = 0.0001，曼-惠特尼检验）。
结论
开发了一种新的简单HPLC方法，用于测定无共洗脱蛋白的尿白蛋白。我们的数据表明，HPLC方法具有高度特异性，且比免疫比浊法更灵敏。
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