HPLC法测定人尿糖比值

Determination of human urinary sugars ratio by HPLC

目的采用HPLC法测定尿液中乳果糖和甘露醇的比值(L/M),以评估健康受试者和手术患者的肠通透性。方法尿样经乙腈沉淀蛋白,离子交换树脂除离子,在Kromasile氨基柱(250 mm×4.6 mm,5μm)上用示差检测器测定尿液中乳果糖和甘露醇的浓度。流动相为乙腈-水(70:30,V/V),流速1 mL·mln^(-1)。结果乳果糖和甘露醇的线性范围为0.1～1.25 g·L^(-1) (r_乳=0.999 2,r_甘=0.999 5),回收率分别为(94.8±2.5)%和(99.1±2.0)%(n=5),日内和日间RSD分别为2.07%～3.45%和4.05%～7.89%、3.44%～5.03%和4.80%～7.37%,最低检出限为0.05 g·L^(-1),健康受试者和手术患者6h的L/M值分别为(0.064±0.024)和(0.132±0.066)。结论该方法可用于评估手术患者的肠通透性改变,但灵敏度不高。

AIM To determine the lactulose and mannitol excretion ratio in urine by HPLC for estimating intestinal permeability of normal subjects and surgical patients. METHODS A human urine was precipitated with aeetonitrile, deionized with ion-exchange resin. The urinary concentrations of lactulose and mannitol were determined by a refractive index detector .A Krornasile NH2 column （250 mm × 4.6 mm,5μm） was used with a mobile phase of acetonitrile-water （70：30, V/V） at a flow rate of 1 mL·min^-1. RESULTS The linear range was0.1 to 1.25 g·L^-1;r was0.9992 for lactulose and 0.999 5 for mannitol, respectively; the average recovery was （94.8 ±2.5） % and （99.1 ± 2.0） % （ n = 5）, for lactulose and mannitol, respectively; intra-day and inter-day precision was from 2.07% to 3.45% and from 4.05% to 7.89%, respectively, for lactulose and from 3.44% to 5.03% and from 4.80% to 7.37%, for mannitol; and the lowest detection limitation for each was 0.05 g· L^- 1. Urinary excretion ratio of lactulose/marmitol of 6 h for normat subjects and surgical patients was （0.064 ± 0.024） and （0. 132 ± 0.066）, respectively. CONCLUSION This method can be used to evaluate the intestinal permeability of surgical patients with limited sensitivity.