不同产地川射干中异黄酮成分含量比较研究

目的：分析测定和对比不同产地川射干中射干苷、鸢尾甲苷A、鸢尾甲苷B、野鸢尾苷、鸢尾苷元、鸢尾甲黄素A、鸢尾甲黄素B 7个异黄酮成分，建立快速测定其含量的方法。方法：采用HPLC法测定7个异黄酮成分含量，色谱柱为Kromasil C₁₈柱（150 mm×4.6 mm，5 μm），以甲醇-0.5%醋酸水溶液为流动相进行梯度洗脱，流速1 mL·min^-1，柱温35℃，检测波长266 nm；并以检测出的含量为指标，使用SPSS 19.0软件对15批样品进行聚类分析。结果：射干苷、鸢尾甲苷A、鸢尾甲苷B、野鸢尾苷、鸢尾苷元、鸢尾甲黄素A、鸢尾甲黄素B色谱峰分离良好；进样量分别在0.024 8~0.992、0.003 6~0.144、0.005 9~0.236、0.004 4~0.176、0.016 2~0.648、0.005 5~0.22、0.005 7~0.228 μg范围内线性关系良好；平均加样回收率（n=6）分别为99.7%、97.7%、98.2%、98.2%、98.8%、103.0%、103.6%，RSD分别为0.57%、2.0%、1.3%、1.1%、0.81%、2.2%、2.3%。15批样品中上述7个成分的含量分别为33.27~58.63、0.62~6.87、1.36~9.95、0.79~7.25、5.42~24.65、0.78~4.56、0.94~8.63 mg·g^-1；聚类分析结果显示15批样品可分为三大类，反映不同产地川射干药材成分的区别。结论：所建立的川射干HPLC含量测定方法可用于川射干的质量控制，聚类分析方法也可为川射干种植、栽培和质量评价提供依据。

Objective:To establish a method for simultaneous determination and comparison of 7 constituents(tectoridin,iristectorin A,iristectorin B,iridin,tectorigenin,iristectorigenin A and iristectorigenin B)in Iridis Tectori Rhizoma from different habitats. Methods:The HPLC assay was conducted on a Kromasil-C₁₈ column(150 mm×4.6 mm,5 μm)with methanol-0.5% acetic acid water solution as a mobile phase. The flow rate was 1.0 mL·min^-1,the detection wavelength was 266 nm and the column temperature was 35℃. Hierarchical clustering analysis(HCA) was perfromed to evaluate and classify 15 batches of samples based on the contents of the seven isoflavones using SPSS 19.0 software. Results:Tectoridin,iristectorin A,iristectorin B,iridin,tectorigenin,iristectorigenin A and iristectorigenin B were separated well. The correlation coefficients between concentration and chromatographic peak area of the 7 componts were over 0.999 9 in the range of 0.024 8-0.992,0.003 6-0.144,0.005 9-0.236,0.004 4-0.176,0.016 2-0.648,0.005 5-0.22 and 0.005 7-0.228 μg,respectively. The average recoveries(n=6)were 99.7%,97.7%,98.2%,98.2%,98.8%,103.0% and 103.6%,respectively with RSDs of 0.57%,2.0%,1.3%,1.1%,0.81%,2.2% and 2.3%. The contents of 7 components in the 15 batches of samples were 33.27-58.63 mg·g^-1 for tectoridin,0.62-6.87 mg·g^-1 for iristectorin A,1.36-9.95 mg·g^-1 for iristectorin B,0.79-7.25 mg·g^-1 for iridin,5.42-24.65 mg·g^-1 for tectorigenin,0.78-4.56 mg·g^-1 for iristectorigenin A and 0.94-8.63 mg·g^-1 for iristectorigenin B. 15 batches of samples were divided into three groups by clustering analysis,which reflected their quality characteristics. Conclusion:The established method can be used for the quality control of Iridis Tectori Rhizoma. HCA can provide basis for planting,cultivation and quality evaluation of the herb.