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目的研究RP-HPLC法分离丰城鸡血藤提取物的条件并建立其中4种异黄酮活性成分的测定方法。方法以乙腈-0.1%磷酸为流动相,采用Spherisord ODS柱(250mm×4mm,5μm),梯度洗脱,体积流量0.8mL/min,检测波长260nm,研究流动相组成和梯度变化对丰城鸡血藤提取物分离效果的影响。结果大豆黄素、染料木素、刺芒柄花素和美皂异黄酮的线性范围、相关系数和平均加样回收率分别为6.24~31.2μg/mL、r=0.9999、99.32%;10.00~130.0μg/mL、r=0.9997、102.20%;5.99~125.9μg/mL、r=0.9999、100.08%和6.96~34.8μg/mL、r=0.9993、99.52%。各组分的加样回收率RSD分别为1.79%、1.51%、1.54%和2.01%(n=5)。结论该法可用于丰城鸡血藤提取物中成分的分离及其中4种异黄酮活性成分的测定。
OBJECTIVE: To study the conditions for the separation of extracts of Spatholobus suberectus from Fengcheng by RP-HPLC and establish a method for the determination of four active ingredients of isoflavones. Methods Acetonitrile-0.1% phosphoric acid was used as the mobile phase. The gradient elution was performed on a Spherisord ODS column (250mm×4mm, 5μm). The volume flow rate was 0.8mL/min. The detection wavelength was 260nm. The mobile phase composition and gradient changes were studied in Fengcheng chicken blood. Effect of vine extract on separation effect. Results The linear range, correlation coefficient and mean sample recovery of daidzein, genistein, acanthophylla and U.S. isoflavones were 6.24~31.2μg/mL, r=0.9999, 99.32%, and 10.00~130.0μg, respectively. /mL, r = 0.9997, 102.20%; 5.99 ~ 125.9 μg/mL, r = 0.9999, 100.08%, and 6.96 ~ 34.8 μg/mL, r = 0.9993, 99.52%. The RSDs for the addition of each component were 1.79%, 1.51%, 1.54%, and 2.01% (n=5), respectively. Conclusion This method can be used for the isolation of components in the extracts of Spatholobus suberectus from Fengcheng and the determination of four active ingredients of isoflavones.