高效阴离子交换色谱法检测酱油中的单糖及双糖

Determination of Monosaccharide and Disaccharide in Soy Sauce by Ion Chromatography

建立了高效阴离子交换色谱-脉冲安培检测法(HPAEC-PAD)测定酱油中阿拉伯糖、半乳糖、葡萄糖、蔗糖、木糖、甘露糖和果糖的方法。采用70%乙醇作为蛋白沉淀剂处理酱油样品,以Dionex CarboPacPA20阴离子交换柱为色谱柱,0.25 mol/L NaOH-水为流动相进行梯度洗脱,考察了色谱柱温度和NaOH浓度对各种糖分离的影响,确定最佳柱温为20℃,NaOH浓度为5.0 mmol/L,7种糖可在20 min内实现基线分离。7种糖的检出限(25μL进样,S/N=3)为2.0～8.0μg/L,线性范围为0.1～80 mg/L,r2大于0.999。酱油样品测定的相对标准偏差为1.4%～4.0%,加标回收率为92%～104%。该方法简便易行,灵敏度高,重现性好,适用于酱油样品中糖组分的分析。

A chromatographic method was developed for the determination of arabinose, galactose, glucose, sucrose, xylose, mannose and fructose in soy sauce sample using high-performance anion-exchange chromatography coupled with pulsed ficiency of the proteins in the soy sauce was amperometric detection（HPAEC -PAD）. The removal efevaluated by using 20% lead acetate, 5% trichloroacetic acid, 15% potassium ferrocyanide or 70% ethanol, respectively. The results indicated that the best sugar recovery was achieved when 70% ethanol was selected as the protein precipitation agent. Effects of column types, column temperature and NaOH concentrations on separation efficiencies of the saccharides were investigated. The optimal conditions were carried out on a Dionex CarboPac PA20 anion-exchange chromatographic column with 0. 25 mol/L 20 ℃. The detection limits（25 μL injection, S/N =3） ranged NaOH -water by gradient eluent at from 2. 0 μg/L to 8.0 μg/L, with a wide linear range of 0. 1 -80 mg/L. The relative standard deviations for the soy sauce sample were between 1.4% and 4. 0% , and the recoveries of standard addition of the saccharides were in the range of 92% - 104%. The method is suitable for the determination of common saccharides in soy sauce samples for with its simplicity, high sensitivity and good reproducibility.