Powell S C, Friedlander E R, Shihabi Z K
J Chromatogr. 1984 Dec 28;317:87-92. doi: 10.1016/s0021-9673(01)91649-8.
Urine and serum myoglobin have been separated on an anion-exchange column, packed by the slurry technique. Urine or serum was injected directly into the column and eluted isocratically with Tris buffer. Freshly prepared myoglobin from human muscle gives two peaks in the chromatogram. Upon storage, one peak slowly decreases while the other increases in size and has the same capacity factor as horse myoglobin. In myoglobinuria, the latter peak is usually detected in urine, while in serum both peaks are detected. For routine assays, horse myoglobin is used as a standard. The minimum detectable level is 2 mg/l. Urine myoglobin from patients with myoglobinuria ranged from 20 to 3000 mg/l, while normal subjects had undetectable levels. Patients with myoglobinuria also had detectable levels of myoglobin in their serum. Urine myoglobin was found to be unstable; it should be analyzed immediately. Although the present method is not sensitive enough to detect myoglobin in the urine of normal subjects, it is clinically useful for confirming and determining myoglobin in patients with myoglobinuria. It has the advantage of speed and simplicity. Using more sensitive detectors would enhance the usefulness of this method.
尿液和血清中的肌红蛋白已通过匀浆技术填充的阴离子交换柱进行分离。将尿液或血清直接注入柱中，并用Tris缓冲液等度洗脱。从人体肌肉中新鲜制备的肌红蛋白在色谱图上呈现两个峰。储存后，一个峰缓慢下降，而另一个峰的大小增加，并且具有与马肌红蛋白相同的容量因子。在肌红蛋白尿中，通常在尿液中检测到后一个峰，而在血清中则检测到两个峰。对于常规检测，使用马肌红蛋白作为标准品。最低检测水平为2mg/L。肌红蛋白尿患者的尿肌红蛋白范围为20至3000mg/L，而正常受试者的尿肌红蛋白水平无法检测到。肌红蛋白尿患者的血清中也可检测到肌红蛋白水平。发现尿肌红蛋白不稳定；应立即进行分析。虽然目前的方法不够灵敏，无法检测正常受试者尿液中的肌红蛋白，但对于确诊和测定肌红蛋白尿患者的肌红蛋白具有临床实用性。它具有速度快和操作简单的优点。使用更灵敏的检测器将提高该方法的实用性。
