测定食物中5种类黄酮物质的高效液相色谱法的改进

Improved HPLC method to determinate 5 types of flavonoids in plant foods

目的优化、完善分析植物性食物中5种类黄酮成分的方法。方法采用Nova-Pak C18(3.9 mm×150mm,4μm)色谱柱;流动相为0.1%三氟乙酸(A)-5%乙腈甲醇溶液(B),梯度洗脱(0～5min,90%A→60%A;5～18 min,60%A→40%A;18～21 min,40%A→10%A;21～25 min,10%A→10%A;25～30 min,10%A→90%A),流速1 ml/min;检测波长365 nm,柱温35℃。结果杨梅黄酮、槲皮素、玉米黄酮、坎二菲醇、芹菜配基5种类黄酮在0.156～20μg/ml浓度范围内与色谱峰高呈良好的线性关系,相关系数(r)为0.9995～0.9999;检测限为0.0195μg/ml(S/N≥3);低、中、高浓度的平均加标回收率(n=3)为88.5%～105.6%,相对标准偏差(RSD)均小于6.32%。结论该方法灵敏、准确,具有良好的重复性和稳定性,可用于植物性食物中5种类黄酮含量的检测。

Objective To improve and optimize the method for the determination of 5 types of flavonoids in plant foods simultaneously.Methods The separation was performed on a Waters Nova-park C18 column（3.9 mm×150 mm,4 μm）;The mobile phase was 0.1% TFA（A）-5% acetonitrile methanol（B）with a gradient elution（0~5 min,90%A→60%A;5~18 min,60%A→40%A;18~21 min,40%A→10%A;21~25 min,10%A→10%A;25~30 min,10%A→90%A）at a flow rate of 1 ml/min;The column temperature was set at 35℃,and the detective wavelength was 365 nm.Results The linear ranges were 0.156~20.0 μg/ml for myricetin,quercetin,luteolin,kaempferol and apigenin.The correlation coefficients（r）were 0.9995~0.9999.The limits of detection was 0.0195 μg/ml（S/N≥3）.The average recoveries（n=3）of 5 types of flavonoids were 88.5 % to 105.6 % with relative standard deviation below 6.32 %.Conclusion This improved method is sensitive,accurate with good repeatability and stability,and can be used for the determination of these 5 types of flavonoids in plant foods.