目的建立高效液相色谱法(HPLC)同时测定纳豆中6种大豆异黄酮含量测定方法。方法通过考察不同的预处理方法,用正交试验设计确定最佳提取条件,优化色谱条件,建立HPLC的测定方法。色谱柱:Agilent Eclipse plus-C18(4.6mm×150mm,5μm);流动相:甲醇+0.5%乙酸梯度洗脱;流速1.0mL/min;检测波长254nm;柱温40℃。结果大豆苷、大豆黄苷、染料木苷、大豆素、大豆黄素和染料木素分别在8.2~82.0、8.3~83.0、8.0~80.0、8.2~82.0、8.6~86.0和8.0~80.0μg/mL范围内线性关系良好;检出限分别为0.037、0.034、0.028、0.019、0.026和0.025μg/mL;回收率95.5%~102.8%;测得纳豆中含有大豆苷1.35mg/g、大豆黄苷0.22mg/g、染料木苷0.96mg/g、大豆素0.17mg/g、大豆黄素0.12mg/g和染料木素0.25mg/g。结论 HPLC法快捷简便、准确可靠、灵敏度高,符合日常检测要求,为纳豆保健功能开发奠定基础。
Objective To develop a HPLC method for the detection of six kinds of soybean isoflavones in natto.Methods By examining the different pretreatment methods and using orthogonal experiment,the optimum extraction conditions were determined.A HPLC method was developed for the detection of soybean isoflavones in natto.The agilent eclipse plus-C18column(5μm,250 mm ×4.6mm)was used with the mobile phase consisting of methanol+ 0.5% acetic acid gradient elution.The detection wavelength was 254 nm.The flow rate was 1.0mL/min and the column temperature was 40 ℃.Results The linear range was 8.2-82.0μg/mL,8.3-83.0μg/mL,8.0-80.0μg/mL,8.2-82.0μg/mL,8.6-86.0μg/mL and 8.0-80.0μg/mL respectively.The detection limit was 0.037μg/mL,0.034μg/mL,0.028μg/mL,0.019μg/mL,0.026μg/mL and 0.025μg/mL respectively.The mean recovery was 95.5%-102.8%.The measured content of Daidzin,Glycitin,Genistin,Daidzein,Glycitein,Genistein was 1.35mg/g,0.22mg/g,0.96mg/g,0.17mg/g,0.12mg/g and 0.25mg/g respectively.Conclusion The HPLC method is simple,rapid,accurate and suitable for daily detecting requirement for further research and development of health function of natto.