大豆异黄酮的提取优化与测定

试验优化了大豆异黄酮的提取方法,建立了一套适用于实验室提取、测定异黄酮含量的高效液相色谱法(HPLC)标准体系。优化后建立的提取条件为:脱脂大豆样品粉末,加入体积浓度为80%的色谱级甲醇10 mL,室温下放置2 h,置于超声功率200 W条件下80℃水浴12 h,12 000 r·min~(-1)离心15 min,取上清液经0.45μm微孔滤膜过滤,待测液4℃保存。优化后的HPLC法:流动相选择甲醇-水(体积比30∶70),柱温设置40℃,波长为254 nm,流速为1 mL·min~(-1),进样体积10μL,进样时间26 min,采用峰面积法计算。结果表明:优化的HPLC法提取大豆异黄酮的效率高、精度高,可为实验室内高效提取大豆异黄酮提供技术参考。

Optimization of the Extraction and Determination of Soybean Isoflavones

In this experiment,the extraction method of soybean isoflavones was optimized,and a high performance liquid chromatography (HPLC) standard measurement system for the determination of isoflavones for laboratory extraction were established.The extraction conditions were as follows:Skimmed soy sample powder added 10 mL of chromatographic grade methanol at a volume of 80％,and placed at room temperature for 2 h.After the installation of ultrasonic power 200 W,80℃ water bath for 12 h,centrifuged at 12 000 r· min-1 for 15 min,taken the supernatant by 0.45 μm microporous membrane filtration,and to be stored at 4℃.The optimal HPLC method was as follows:The mobile phase was methanol-water (volume ratio 30∶70),column temperature set at 40℃,the wave length was 254 nm,the flow rate was 1 mL· min-1,the sample volume was 10 μL,set injection time for 26 min,and calculated by peak area method.The results showed that the optimized HPLC method was efficient and accurate in extracting soybean isoflavones.It could provide technical reference for efficient extraction of soy isoflavones in laboratory.