反相高效液相色谱法测定大豆籽粒中异黄酮含量的方法研究

Determination of Isoflavone Content in Soybean by Reverse Phase High Performance Liquid Chromatography

大豆异黄酮是一类具有特殊生理活性物质。研究并建立了应用反相高效液相色谱法测定大豆籽粒中黄豆苷、黄豆黄素苷和染料木苷3种异黄酮成分含量的检测方法。同时，运用统计学原理对建立的大豆异黄酮测定方法进行了系统、全面的分析和评价。结果表明：黄豆苷、黄豆黄素苷和染料木苷线性范围分别为1．25～20．0μg·mL^-1、0．625～10．0μg·mL^-1和1．25—20．0μg·mL^-1；回收率范围为97．1％-99．5％；重复性测定的相对相差均低于5％；不同实验室对黄豆苷、黄豆黄素苷和染料木苷测定的相对标准偏差分别为1．15％-3．42％、2．44％-4．84％和1．43％～3．06％。上述结果说明该方法的线性、准确度、精密度、同一实验室重复性和多家实验室的再现性评价结果优良，适合作为国家农业行业标准的制定和实施。

Isoflavone is a group of special biofunctional substance. In this paper, a method determining isoflavone including Daidzin, Glycitin and Genistin by Reverse Phase High Performance Liquid Chromatography （RP-HPLC） was investigated and erected. Meanwhile ,the isoflavone determination method was analysed as well as evaluated wholly and systematically by statistical technique. The results showed that the linear range of Daidzin, Glycitin and Genistin was 1.25 -20.0μg·mL^-1, 0.625 - 10.0μg·mL^-1 and 1.25 - 20.0 μg·mL^-1, respectively ; the recovery range was 97.1% - 99.5 % ; the relative deflation of repeatable test was below 5% ;the relative standard deviation of Daidzin, Glycitin and Genistin among different labs was 1.15% -3.42% ,2.44% -4.84% and 1.43% -3.06%, respectively. It showed from the above analysis that this method possessed good linearity, good quality of accuracy, precision, repeatability in one lab and reproducibility among labs. Therefore,this method was proved to be suitable for erecting and implementing a national agricultural standard.