Tajima S, Hashiba M, Suzuki T, Akanuma H, Yabuuchi M
Takasaki Research Laboratories, Nippon Kayaku Co. Ltd., Gumma, Japan.
Biomed Chromatogr. 1993 Jan-Feb;7(1):41-4. doi: 10.1002/bmc.1130070111.
A simple high performance liquid chromatographic method combined with an enzyme sensor has been developed to measure 1,5-anhydroglucitol in urine. The enzyme sensor consists of a hydrogen peroxide electrode and a chitosan membrane of an immobilized pyranose oxidase. As the system does not resist interfering substances, urine samples are first purified by passing them through a two-layer column packed with (1) strongly basic anion (OH- form, the upper layer) and (2) strongly acidic cation (H+ form, the lower layer) exchange resins. 1,5-Anhydroglucitol is efficiently recovered in the flow-through fraction of the column. In this system, the minimum detectable concentration of 1,5-anhydroglucitol is 0.1 mg/L, and the measurable range extends from 0.1 to 60 mg/L. The coefficient of variation values of the within-day and day-to-day precisions are 3.0-6.5% and and 4.4-6.7% respectively, and there is good agreement between the results measured by our method and those obtained by the gas-liquid chromatographic/mass spectrometric method (r = 0.994). The method we have described here has been successfully used to elucidate a mechanism for the reducing 1,5-anhydroglucitol level in the serum and plasma of patients.
已开发出一种结合酶传感器的简单高效液相色谱法来测定尿液中的1,5-脱水葡萄糖醇。该酶传感器由过氧化氢电极和固定化吡喃糖氧化酶的壳聚糖膜组成。由于该系统不能抵抗干扰物质，尿液样品首先通过装有（1）强碱性阴离子（OH-型，上层）和（2）强酸性阳离子（H+型，下层）交换树脂的两层柱进行纯化。1,5-脱水葡萄糖醇在柱的流出部分中被有效地回收。在该系统中，1,5-脱水葡萄糖醇的最低可检测浓度为0.1mg/L，可测量范围为0.1至60mg/L。日内和日间精密度的变异系数值分别为3.0-6.5%和4.4-6.7%，并且我们的方法所测结果与气液色谱/质谱法所得结果之间具有良好的一致性（r = 0.994）。我们在此描述的方法已成功用于阐明患者血清和血浆中1,5-脱水葡萄糖醇水平降低的机制。
