超高效液相色谱法同时测定保健品中4种大豆异黄酮

Simultaneous determination of 4 kinds of soybean isoflavones in health products by ultra performance liquid chromatography

目的建立超高效液相色谱法(ultraperformance liquid chromatography,UPLC)同时测定保健品中大豆苷(daidzin)、染料木苷(genistin)、染料木素(daidzein)和大豆素(genistein)4种大豆异黄酮的分析方法。方法样品用80%甲醇超声提取后,以甲醇-0.1%磷酸水(V/V)作为流动相梯度洗脱分离,使用二极管阵列检测器(photo-diode array, PDA)于260 nm波长下测定。结果 4种大豆异黄酮在1.0~50 mg/L范围内,标准曲线线性良好,相关系数r2均大于0.999。平均回收率为86.7%~106%,相对标准偏差(relative standard deviation, RSD)为1.31%~6.32%。方法检出限为0.5~1.0mg/kg,方法定量限为1.5~3.0mg/kg。结论该方法回收率高,灵敏度高,且操作简单,快速高效,可应用于实际样品测定。

Objective To establish a method for the determination of 4 soybean isoflavones in health products included daidzin, genistin, daidzein and genistein by ultra performance liquid chromatography(UPLC). Method Samples were ultrasonic extracted by 80% methanol-water, then separated by methanol-0.1% phosphoric acid-water(V/V) as mobile phase with gradient elution, finally detected by photo-diode array detector under the wavelength of 260 nm. Results The standard curve of four soybean isoflavones exhibited a good linearity in the concentration range of 1.0-50 mg/L, while the correlation coefficient(r2) were both greater than 0.999. Moreover, the average recoveries were ranged from 86.7% to 106% with the relative standard deviations were ranged from 1.31% to 6.32%. The limits of detection and the limits of quantitation were 0.5-1.0 mg/kg and 1.5-3.0 mg/kg respectively. Conclusion This method has high recovery rate and high sensitivity. It is simple operation, fast and efficient that can be applied to actual determination.