高效液相色谱法同时测定粮食中6种大豆异黄酮

Determination of 6 kinds of isoflavones in food crops by high performance liquid chromatography

中文关键词:  高效液相色谱法  大豆异黄酮  粮食

英文关键词:high performance liquid chromatography  isoflavones  food crops

中文摘要:

      目的 建立高效液相色谱法(high performance liquid chromatography, HPLC)测定粮食中6种大豆异黄酮(大豆苷、大豆黄苷、染料木苷、大豆素、大豆黄素和染料木素)含量的分析方法。方法 准确称取一定量粉碎混匀后的样品, 经80%甲醇提取后取上清液, 过0.22 μm有机相滤膜上机。采用Thermo Syncronis C18柱(250 mm×4.6 mm, 5 μm), 以0.5%甲酸水溶液(A)、乙腈(B)和甲醇(C)作为流动相进行梯度洗脱, 流速0.8 mL/min, 柱温为35 ℃, 于紫外检测器波长260 nm处检测, 大豆异黄酮各组分含量以外标法进行定量。结果 本方法在30 min内完成6种大豆异黄酮的分离分析, 大豆异黄酮各组分浓度在0.2~50 μg/mL范围内呈良好的线性关系(r>0.999), 平均加标回收率为96.9%~107.8%, 相对标准偏差(relative standard deviation, RSD)为0.6%~5.0%, 检出限为0.03~0.1 μg/mL, 定量限为0.1~0.3 μg/mL。结论 建立的方法具有较高的灵敏度和重复性, 能满足不同粮食种类中大豆异黄酮含量测定。

英文摘要:

      Objective To establish a method for simultaneous determination of 6 kinds of isoflavones (daidzin, glycitin, genistin, daidzein, glycitein and genistein) in food crops by high performance liquid chromatography (HPLC). Methods A certain amount of crushed mixed samples were weighed accurately, followed by extracting with 80% methanol, then the supernatant was filtered through 0.22 μm membrane microfilter for analysis. The samples were separated with a Thermo Syncronis C18 column (250 mm×4.6 mm, 5 μm) at the flow rate of 0.8 mL/min by gradient elution using 0.5% formic acid water (solvent A), acetonitrile (solvent B) and methanol (solvent C). The temperature of the column oven was maintained at 35 ℃ and the signal was detected with ultraviolet detector in 260 nm, and the contents of 6 kinds of isoflavones were quantitatively analyzed by external standard method. Results The 6 kinds of isoflavones were separated within 30 min, each compound showed a good linear relationship between peak areas and concentrations in the concentration range of 0.2~50 μg/mL (r>0.999). The recoveries of 6 kinds of isoflavones in food crops were 96.9%~107.8% with relative standard deviations (RSD) of 0.6%~5.0%. The limits of detection (LODs) were 0.03~0.1 μg/mL and the limit of quantification (LOQs) were 0.1~0.3 μg/mL. Conclusion The established method exhibits high sensitivity and repeatability, which can satisfy the determination of isoflavones in different food crops.