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目的:建立同时测定葛花中大豆苷、鸢尾苷、大豆苷元和鸢尾苷元4种异黄酮含量的快速分析方法。方法:采用Agilent C18色谱柱(250 mm×4.6 mm,5μm),以甲醇-乙腈-水(2∶1∶2)为流动相,流速为1.0 ml·min-1;检测波长为264 nm,柱温为25℃,进样量为20μl。结果:4种异黄酮的保留时间分别3.4,3.8,5.7,7.2 min,一次分析可在10 min内完成。其线性范围分别为0.784~78.440μg·ml-1(大豆苷)、2.000~200.000μg·ml-1(鸢尾苷)、0.800~80.020μg·ml-1(大豆苷元和鸢尾苷元);r分别为0.999 9,0.999 8,0.999 7,0.999 9,平均回收率分别为100.54%(RSD=1.66%,n=6)、100.03%(RSD=1.00%,n=6)、99.48%(RSD=1.76%,n=6)和100.92%(RSD=2.26%,n=6)。结论:本方法简便易行,快速准确,重复性好,可以为葛花异黄酮的研发提供方便快捷的检测方法。
OBJECTIVE: To establish a rapid method for the simultaneous determination of daidzin, tectorigenin, daidzein and tectorigenin in four kinds of isoflavones. METHODS: The mobile phase consisted of methanol-acetonitrile-water (2:1:2) with a flow rate of 1.0 ml · min-1 on an Agilent C18 column (250 mm × 4.6 mm, 5 μm) The temperature was 25 ° C and the injection volume was 20 μl. Results: The retention times of four isoflavones were 3.4, 3.8, 5.7 and 7.2 minutes, respectively. The first analysis could be completed within 10 minutes. The linear range was 0.784 ~ 78.440μg · ml-1 (daidzin), 2.000 ~ 200.000μg · ml-1 (tectorigen), 0.800 ~ 80.020μg · ml-1 (daidzein and tectorigenin) (RSD = 1.66%, n = 6), 100.03% (RSD = 1.00%, n = 6) and 99.48% (RSD = 1.76%, n = 6) and 100.92% (RSD = 2.26%, n = 6). Conclusion: The method is simple, rapid, accurate and reproducible, which can provide a convenient and rapid method for the determination of genistein.