Yamamoto K, Okano K, Yamamoto K, Yamashiro Y, Hattori Y, Ohba Y, Fujii Y
School of the Allied Health Science, Yamaguchi University, Ube.
Rinsho Byori. 1994 Feb;42(2):200-4.
In vitro globin chain synthesis ratios (beta/alpha) were determined using high performance liquid chromatography (HPLC) in twenty-five alpha or beta-thalassemiaes as well as six normal controls. Among thalassemiaes nineteen were Japanese and six Korean. Two kinds of cation exchange columns.; Mono S, HR 5/5 by Pharmacia and TSK gel by Tohso were compared. Hemolysates (10%), in stead of purified globin peptides, were directly subjected to the both columns, and 3H-labeled globin chain in vitro synthesized were separated on them. The Tohso column gave better resolution of globin peptides, resulting in sharp elution profiles. The HPLC system took about two hours for an analysis of a sample. In contrast, the conventional carboxymethyl cellulose (CMC) chromatography by Clegg need larger sample amounts for globin extraction and twenty-four hours to be completed. The HPLC is evidently simple and rapid. The globin chain ratios thus obtained by HPLC were 1.04-1.08, 0.23-0.89, 1.22-1.62 and 2.05-4.29 for normals, thirteen beta-thalassemia minors, seven alpha-thalassemia traits and three Hb H diseases respectively.
采用高效液相色谱法（HPLC）测定了25例α或β地中海贫血患者以及6例正常对照者的体外珠蛋白链合成比率（β/α）。在地中海贫血患者中，19例为日本人，6例为韩国人。比较了两种阳离子交换柱，即Pharmacia公司的Mono S、HR 5/5柱和Tohso公司的TSK凝胶柱。直接将10%的溶血产物而非纯化的珠蛋白肽加样到两根柱上，分离体外合成的3H标记珠蛋白链。Tohso柱对珠蛋白肽的分离效果更好，洗脱图谱更尖锐。HPLC系统分析一个样品约需两小时。相比之下，Clegg的传统羧甲基纤维素（CMC）色谱法提取珠蛋白所需样品量更大，且需24小时才能完成。HPLC显然简便快捷。通过HPLC获得的正常对照者以及13例轻型β地中海贫血、7例α地中海贫血特征患者和3例血红蛋白H病患者的珠蛋白链比率分别为1.04 - 1.08、0.23 - 0.89、1.22 - 1.62和2.05 - 4.29。
