HPLC法快速测定葛花提取物中4种异黄酮类化合物的含量

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。

Quick Detection of Four Isoflavone Compounds from Extracts of Pueraria Lobata Flowers by HPLC

Objective： To establish an HPLC method for the determination of four kinds of isoflavone compounds including daidzin,tectoridin,daidzein and tectorigenin in the extracts of Pueraria lobata flowers. Methods： The separation was performed on an Agilent C18column（ 250 mm × 4. 6 mm,5 μm） using a mobile phase of methanol-acetonitrile-water（ 2∶ 1∶ 2）. The flow rate was 1. 0 ml·min- 1,and the detection wavelength was 264 nm. The column temperature was 25℃ and the sample size was 20 μl. Results： The detection could be accomplished within 10 minutes with good separation and specificity of four isoflavone compounds with the retention time of 3. 4,3. 8,5. 7 and 7. 2 min,respectively. The linear range was 0. 784-78. 440 μg·ml- 1（ daidzin）,2.000-200.000 μg·ml- 1（ tectoridin） and 0.800-80.020 μg·ml- 1（ daidzein and tectorigenin）,and the relative coefficient was 0.999 9,0.999 8,0. 999 7 and 0. 999 9,respectively. The average recovery was 100. 54%（ RSD = 1. 66%,n = 6）,100. 03%（ RSD = 1. 00%,n = 6）,99. 48%（ RSD = 1. 76%,n = 6） and 100. 92%（ RSD = 2. 26%,n = 6）,respectively. Conclusion： The method is simple,rapid and accurate with good repeatability,which can be used in the rapid determination of isoflavone compounds in the flowers of Pueraria lobata.