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利用超声波技术,以大豆异黄酮主要组分染料木素为对照,根据单因素试验和正交试验结果建立三波长比色法和紫外分光光度法,确定大豆异黄酮最佳提取工艺:乙醇浓度70%,料液比1∶25,提取温度50℃,提取时间5 h,提取2次。比较新建立方法与之前建立HPLC法的精密度和准确度,结果依次为:高效液相色谱法>三波长比色法>紫外分光光度法,其中HPLC法灵敏度高,适于大豆及其制品中异黄酮含量的精确测定,三波长比色法和紫外分光光度法适于异黄酮含量批量、快速测定。采用HPLC法测得东北地区品种异黄酮含量分布范围为1.0404～4.4344μg.mg-1,平均值2.9174μg.mg-1,不同地区大豆品种异黄酮总量分布规律为:黑龙江>吉林>其它地区(主要是辽宁省和内蒙古东部地区),筛选得到大于4.0μg.mg-1高异黄酮品种12份,小于1.0μg.mg-1的低异黄酮品种3份。
Using ultrasonic technology, the main component of soybean isoflavones genistein as a control, according to the single factor test and orthogonal test results to establish three-wavelength colorimetry and UV spectrophotometry to determine the optimum extraction of soy isoflavones: ethanol concentration 70 %, Solid-liquid ratio 1:25, extraction temperature 50 ℃, extraction time 5 h, extraction 2 times. Compared with the established method and HPLC method, the precision and accuracy of the method were compared. The results were as follows: high performance liquid chromatography> three-wavelength colorimetric method> UV spectrophotometry. Among them, the HPLC method was sensitive and suitable for soybean and its products Accurate determination of isoflavone content, three wavelength colorimetry and UV spectrophotometry for isoflavone content batch, rapid determination. The content of isoflavones in Northeast China ranged from 1.0404 to 4.4344μg.mg-1 and the average value was 2.9174μg.mg-1 by HPLC. The distribution of total isoflavones in different regions was Heilongjiang> Jilin> other regions (Mainly in Liaoning Province and eastern Inner Mongolia), screening 12 parts of isoflavone more than 4.0μg.mg-1 and 3 parts of low-isoflavone less than 1.0μg.mg-1.