HPLC-DAD法同时测定中药金钱草中6种活性成分的含量

Simultaneous Determination of Six Active Constituents in Herba Lysimachiae by HPLC-DAD

目的:建立HPLC-DAD法同时测定中药金钱草中绿原酸、槲皮素、山奈酚、山奈酚-3-O-β-D-葡萄糖苷、芦丁、蒙花苷6种活性成分的含量。方法:Diamonsil C_(18)(200 mm×4.6 mm,5μm)色谱柱,流动相为甲醇(A)-0.3%磷酸水溶液(B),梯度洗脱,流速:1.0 ml·min^(-1),检测波长:360 nm,柱温:30℃。结果:绿原酸、槲皮素、山奈酚、山奈酚-3-O-β-D-葡萄糖苷、芦丁、蒙花苷的质量分别在0.031~0.624μg(r=0.999 6)、0.019~0.386μg(r=0.999 6)、0.027~0.534μg(r=0.999 7)、0.032~0.644μg(r=0.999 8)、0.019~0.376μg(r=0.999 8)、0.017~0.340μg(r=0.999 7)内与峰面积呈良好的线性关系。绿原酸、槲皮素、山奈酚、山奈酚-3-O-β-D-葡萄糖苷、芦丁、蒙花苷的平均加样回收率分别为97.35%、97.98%、98.43%、99.57%、98.02%和97.92%,RSD分别为0.87%、1.53%、1.46%、1.24%、1.10%和1.41%(n=6)。结论:该方法简便、准确、重复性好,可用于金钱草药材多成分的质量控制研究。

Objective： To develop an HPLC-DAD method for the simultaneous determination of six active constituents of chlorogenic acid,quercetin,kaempferol,kaempferol-3-O-β-D-glucoside,rutin and linarin in Herba Lysimachiae. Methods： The chromatographic separation was performed on a Diamonsil C_（18）column（ 200 mm × 4. 6 mm,5 μm） with the mobile phase of 0. 3% phosphoric acid-methanol with gradient elution at the flow rate of 1. 0 ml·min^（-1）. The detection wavelength was set at 360 nm,and the column temperature was maintained at 30℃. Results： Chlorogenic acid,quercetin,kaempferol,kaempferol-3-O-β-D-glucoside,rutin and linarin was linear within the range of 0. 031-0. 624 μg（ r = 0. 999 6）,0. 019-0. 386 μg（ r = 0. 999 6）,0. 027-0. 534 μg（ r =0. 999 7）,0. 032-0. 644 μg（ r = 0. 999 8）,0. 019-0. 376 μg（ r = 0. 999 8） and 0. 017-0. 340 μg（ r = 0. 999 7）,respectively. The average recovery was 97. 35%,97. 98%,98. 43%,99. 57%,98. 02% and 97. 92% with RSD of 0. 87%,1. 53%,1. 46%,1. 24%,1. 10% and 1. 41%（ n = 6）,respectively. Conclusion： The method is convenient,accurate and reproducible in the quality control of multi-components in Herba Lysimachiae.