高效液相色谱法测定蛋白粉中异麦芽酮糖含量

Determination of isomaltulose content in protein powder by high performance liquid chromatography

中文关键词:  高效液相色谱法(HPLC)  异麦芽酮糖  食品  HILIC

英文关键词:high performance liquid chromatography  isomaltulose  protein powder  hydrophilic interaction liquid chromatography column

中文摘要:

      目的 建立高效液相色谱法(high performance liquid chromatography, HPLC)测定蛋白粉中异麦芽酮糖含量。方法 样品经50%乙腈超声萃取, 离心后取上清液, 经 0.45 μm微孔滤膜过滤后用亲水作用色谱(hydrophilic interaction liquid chromatography column, HILIC)分离, 高效液相色谱仪-示差检测器分析。选用色谱柱Waters XBridge HILIC (4.6 mm×250 mm, 5 μm), 流动相: 乙腈:水=85:15 (V:V); 检测器温度为40 ℃; 流速为1.0 mL/min; 进样体积为5 μL; 柱温为40 ℃; 运行时间为10 min。结果 异麦芽酮糖在1.96~9.80 mg/mL范围内线性关系良好, 相关系数(r2)为0.9999, 回收率均大于90% (n=9), 相对标准偏差为0.65%。本方法检出限为0.01 g/100 g, 定量限为0.04 g/100 g。结论 本方法前处理方法简单, 检测灵敏度高, 稳定性好, 适用于蛋白粉样品中异麦芽酮糖含量的检测。样品使用50%乙腈水溶解样品能较好地除去部分杂质, 同时采用Xbridge HILIC柱, 有较好的极性保留, 检测时间短, 分离效果好, 可以避免其他糖成分带来的干扰。

英文摘要:

      Objective To establish a method for the determination of isomaltulose content in protein powder by high performance liquid chromatography (HPLC). Methods The sample was extracted by 50% acetonitrile ultrasonic extraction, centrifuged, and the supernatant was taken. It was filtered through a 0.45 μm microporous membrane and separated by hydrophilic interaction liquid chromatography column (HILIC). The sample was analyzed by high-performance liquid chromatography differential detector. Select the Waters XBridge HILIC chromatographic column (4.6 mm×250 mm, 5 μm), mobile phase: Acetonitrile:water=85:15 (V:V); the detector temperature was 40 ℃; the flow rate was 1.0 mL/min; the injection volume was 5 μL; the column temperature was 40 ℃; the running time was 10 minutes. Results The linear relationship of isomaltulose was good within the range of 1.96–9.80 mg/mL, with a correlation coefficient of 0.9999 and recovery rates greater than 90% (n=9). The relative standard deviation value was 0.65%. The limit of detection of this method was 0.01 g/100 g, and the limit of quantification was 0.04 g/100 g. Conclusion This method has a simple pre-processing method, high detection sensitivity, and good stability, and is suitable for detecting the content of isomaltulose in protein powder samples. Dissolving the sample in 50% acetonitrile water can effectively remove some impurities, while using an Xbridge HILIC column has good polarity retention, short detection time, good separation effect, and can avoid interference from other sugar components.