Akanuma H, Yamanouchi T, Ono H, Nomura K, Akanuma Y
J Biochem. 1985 Feb;97(2):579-88. doi: 10.1093/oxfordjournals.jbchem.a135093.
This report describes an application of liquid chromatography to the determination of sorbitol in red blood cells. The chromatograph employed in the present study was made up of sub- and main-separation systems and a detector portion. The sub-separation system was for concentration of polyols and involved two small columns, each containing the same anion exchange resin. The first was a tiny column which, in borate form, served as the concentrator of polyols and sugars charged in a large volume, while the second, in acetate form, separated the carbohydrates from the borate. The main system was for the fine separation of each carbohydrate and employed cation exchange columns. The detector part utilized a flow fluorometric method comprising two successive reactions: periodate oxidation followed by the Hantzsch reaction. The resulting whole chromatographic system was applied to the determination of sorbitol in red blood cells obtained from normal rats and rats made diabetic by the administration of streptozotocin; a part of the latter group had also received an aldose reductase inhibitor. Our results supported the concepts that a prolonged duration of high blood glucose level induces an elevated level of sorbitol inside red blood cells and that aldose reductase inhibitors are effective in reducing this level.
本报告描述了液相色谱法在测定红细胞中山梨醇含量方面的应用。本研究中使用的色谱仪由子分离系统、主分离系统和检测部分组成。子分离系统用于多元醇的浓缩，包含两根小柱，每根小柱都装有相同的阴离子交换树脂。第一根是微型柱，以硼酸盐形式作为大量注入的多元醇和糖的浓缩器，而第二根以醋酸盐形式，将碳水化合物与硼酸盐分离。主系统用于每种碳水化合物的精细分离，采用阳离子交换柱。检测部分采用流动荧光法，包括两个连续反应：高碘酸盐氧化，然后是汉茨希反应。所得的整个色谱系统应用于测定从正常大鼠和通过注射链脲佐菌素诱导糖尿病的大鼠获得的红细胞中的山梨醇；后一组中的一部分大鼠还接受了醛糖还原酶抑制剂。我们的结果支持了以下观点：高血糖水平持续时间延长会导致红细胞内山梨醇水平升高，且醛糖还原酶抑制剂可有效降低该水平。
