HPLC法同时测定黄花獐牙菜根中8种成分

Simultaneous determination of eight components in root of Swertia kingii by HPLC

目的建立同时测定黄花獐牙菜Swertia kingii根中獐牙菜苷、龙胆苦苷、异荭草苷、1,3,7,8-四羟基酮-1-O-β-D-葡萄糖、当药醇苷、1,3,7,8-四羟基酮、去甲基雏菊叶龙胆酮和1,5,8-三羟基-3-甲氧基酮8种成分的HPLC方法。方法采用Agilent Zorbax ODS(250 mm×4.6 mm,5μm)色谱柱,乙腈-0.5%磷酸水梯度洗脱,体积流量为1.0 mL/min,检测波长254 nm,柱温30℃。结果獐牙菜苷、龙胆苦苷、异荭草苷、1,3,7,8-四羟基酮-1-O-β-D-葡萄糖、当药醇苷、1,3,7,8-四羟基酮、去甲基雏菊叶龙胆酮和1,5,8-三羟基-3-甲氧基酮分别在3.84~96.00μg/mL(r=0.999 6)、3.36~84.00μg/mL(r=0.999 2)、5.92~148.00μg/mL(r=0.999 8)、4.81~118.00μg/mL(r=0.999 2)、4.32~108.00μg/mL(r=0.999 3)、4.16~104.00μg/mL(r=0.999 2)、5.12~128.00μg/mL(r=0.999 4)、4.80~120.00μg/mL(r=0.999 6)与峰面积积分值呈良好的线性关系;平均回收率分别为99.59%、98.95%、98.61%、99.63%、99.31%、99.50%、99.80%、98.50%,RSD分别为0.99%、3.37%、1.87%、1.93%、1.21%、1.62%、0.54%、1.87%。结论所建方法简便、高效、准确,重复性好,可用于对黄花獐牙菜的质量控制,有利于对该植物的综合利用开发。

Objective To establish an HPLC method for simultaneously determining eight components, such as sweroside, gentiopicroside, isoorientin, 1,3,7,8-tetrahydroxy-xanthone-1-O-β-D-glucopyranosyl, swertianolin, 1,3,7,8-tetrahydroxy-xanthone, demethylbellidifolin, and 1,5,8-trihydrox-3-methoxyxanthone in the root of Swertia kingii. Methods Chromatographic analysis was achieved on an Agilent Zorbax ODS column（250 mm × 4.6 mm, 5 μm） by gradient elution of acetonitrile-0.5% phosphoric acid in water at 30 ℃. The flow rate was 1.0 mL/min and the detection wavelength was 254 nm. Results The calibration curves of all the eight constituents showed good linearity in a relatively wide concentration range. The linear ranges of swertiamarin, gentiopicroside, isoorientin, 1,3,7,8-tetrahydroxy-xanthone-1-O-β-D-glucopyranosyl, swertianolin, 1,3,7,8-tetrahydroxy-xanthone, demethylbellidifolin, and 1,5,8-trihydrox-3-methoxyxanthone were 3.84—96.00 μg/mL（r = 0.999 6）, 3.36—84.00 μg/mL（r = 0.999 2）, 5.92—148.00 μg/mL（r = 0.999 8）, 4.81—118.00 μg/mL（r = 0.999 2）, 4.32—108.00 μg/mL（r = 0.999 3）, 4.16—104.00 μg/mL（r = 0.999 2）, 5.12—128.00 μg/mL（r = 0.999 4）, 4.80—120.00 μg/mL（r = 0.999 6）, respectively. The average recoveries were 99.59%（RSD 0.99%）, 98.95%（RSD 3.37%）, 98.61%（RSD 1.87%）, 99.63%（RSD 1.93%）, 99.31%（RSD 1.21%）, 99.50%（RSD 1.62%）, 99.80%（RSD 0.54%）, and 98.50%（RSD 1.87%）. Conclusion This method is simple, efficient, accurate, and reproducible, and can be used for the determination of eight constituents in root of Swertia kingii. This will promote the comprehensive usage of this plant.