Huisman T H, Henson J B, Wilson J B
J Lab Clin Med. 1983 Aug;102(2):163-73.
A new HPLC procedure is described for the separation and quantitation of minor human Hb variants. The cation exchanger, Synchropak CM 300, is a silica support with a bonded polymeric coating of carboxylic acid residues. The chromatogram is developed with 0.03M Bis-Tris-KCN buffers, pH 6.40, and a Na-acetate gradient increasing from 0 to 0.1125M. As many as 11 minor Hbs (including Hb A2) can be isolated. Some of these have been identified through rechromatography of the minor Hb zones obtained by Bio-Rex-70 chromatography. Quantitation of Hb A1c and Hb A2 is readily accomplished. Hb F0 and an unidentified minor Hb often observed in red cells of alcoholics co-chromatograph with Hb A1c. The method has been applied to blood samples of 10 normal adults, 13 diabetic patients, and nine alcoholic subjects. An excellent correlation exists between the Hb A1c percentages and the levels of Hb A1 determined by microcolumn chromatography. Some other minor Hbs, identified as components 9 and 10, which are (at least in part) Hb A0 with glucose attached to the alpha chains, are present in increased amounts in the blood of diabetic patients and others may be observed in patients who subject themselves to (severe) alcohol abuse. It is suggested that the new procedure is well-suited for detailed studies of the minor Hbs in patients with various abnormalities in carbohydrate metabolism.
本文描述了一种用于分离和定量分析人血红蛋白（Hb）微量变体的高效液相色谱（HPLC）新方法。阳离子交换剂Synchropak CM 300是一种硅胶载体，表面键合有含羧酸残基的聚合物涂层。色谱分离采用0.03M的Bis-Tris-KCN缓冲液（pH 6.40），并用从0到0.1125M递增的醋酸钠梯度洗脱。该方法可分离多达11种微量血红蛋白（包括Hb A2）。其中一些通过对Bio-Rex-70色谱法得到的微量Hb区带进行再色谱分析得以鉴定。Hb A1c和Hb A2的定量分析很容易实现。Hb F0以及在酗酒者红细胞中经常观察到的一种未鉴定的微量Hb与Hb A1c共色谱。该方法已应用于10名正常成年人、13名糖尿病患者和9名酗酒者的血样分析。Hb A1c百分比与微柱色谱法测定的Hb A1水平之间存在良好的相关性。另外一些被鉴定为组分9和10的微量Hb（至少部分是α链上连接有葡萄糖的Hb A0），在糖尿病患者血液中的含量增加，而在酗酒（严重）患者中可能会观察到其他微量Hb。建议该新方法非常适合对碳水化合物代谢异常患者的微量Hb进行详细研究。
