目的 建立测定西藏秦艽花和川西秦艽花中落干酸、6'-O-β-D-葡萄糖基龙胆苦苷、獐芽菜苦苷、龙胆苦苷、獐芽菜苷、异荭草苷、异牡荆黄素7种成分的HPLC方法.方法 采用Agilent Zorbax ODS(250 mm×4.6 mm,5 μm)色谱柱,流动相为乙腈-0.4%醋酸水溶液梯度洗脱,体积流量0.4 mL/min,检测波长254 nm,柱温30 ℃.结果 落干酸、6'-O-β-D-葡萄糖基龙胆苦苷、獐芽菜苦苷、龙胆苦苷、獐芽菜苷、异荭草苷和异牡荆黄素分别在0.228～2.280、0.680～6.800、0.220～2.200、1.476～14.760、0.200～2.000、0.436～4.360、0.276～2.760 μg进样量与峰面积积分值线性关系良好(R²≥0.999 2),落干酸、6'-O-β-D-葡萄糖基龙胆苦苷、獐芽菜苦苷、龙胆苦苷、獐芽菜苷、异荭草苷及异牡荆黄素的平均回收率(n＝9)分别为100.9%、99.4%、101.0%、105.7%、103.1%、98.4%、104.2%.结论 该法简便、准确、专属性强,可用于西藏秦艽花和川西秦艽花中7种成分的测定.

Objective To establish an HPLC method for simultaneously determining seven components, such as loganic acid, 6'-O-β-D-glucopyranosyl gentiopicroside, swertiamarine, gentiopicroside, swertiamarin, isoorientin, and isovitexin, from the flowers of Gentiana tibetica and G. dendrology. Methods Chromatographic analysis was achieved on an Agilent Zorbax ODS column (150 mm × 4.6 mm, 5 μm) by gradient elution of acetonitrile-0.4% acetic acid in water at 30 ℃. The flow rate was 0.4 mL/min and the detection wavelength was 254 nm. Results The calibration curves of all the seven constituents showed good linearity in a relatively wide concentration range. The linear ranges of loganic acid, 6'-O-β-D-glucopyranosyl gentiopicroside, swertiamarine, gentiopicroside, gentiopicroside, isoorientin, and isovitexin were 0.228—2.280, 0.680—6.800, 0.220—2.20, 1.476—14.760, 0.200—2.000, 0.436—4.360, and 0.276—2.760 μg (R²≥ 0.999 2), respectively. The recoveries (n = 9) of loganic acid, 6'-O-β-D-glucopyranosyl gentiopicroside, swertiamarine, gentiopicroside, gentiopicroside, isoorientin, and isovitexin were 100.9%, 99.4%, 101.0%, 105.7%, 103.1%, 98.4%, 104.2%. Conclusion This method is simple, accurate, and specific, and can be used for the determination of seven constituents in the flowers of G. tibetica and G. dendrologi.

西南民族大学2014级研究生“创新型科技项目”(CX2014SZ47)