HPLC波长切换法同时测定粗茎秦艽中6个成分的含量

HPLC with switching wavelength simultaneous determination of six constituents in Gentiana crassicaulis

本实验建立了HPLC波长切换法同时测定云南丽江粗茎秦艽中马钱苷酸、獐牙菜苦苷、龙胆苦苷、獐牙菜苷、异荭草苷、异牡荆苷6种成分的含量,并对各成分含量间的相关性进行分析,为粗茎秦艽药材质量的全面评价提供了科学依据。采用Sepax Gp-C18（150 mm×4.6 mm,5μm）色谱柱,甲醇-0.1%磷酸为流动相体系,梯度洗脱,流速1.0 mL·min-1,柱温25℃,检测波长235 nm（0～51 min）、270 nm（51～57 min）、243 nm（57～65 min）、270 nm（65～80 min）。马钱苷酸、獐牙菜苦苷、龙胆苦苷、獐牙菜苷、异荭草苷、异牡荆苷的量分别在0.01000～10.0000、0.00216～2.1600、0.0480～48.0000、0.0010～1.0000、0.00034～0.3400、0.00026～0.2600μg范围内与色谱峰峰面积呈良好的线性关系,平均回收率为98.52%～99.31%,RSD均小于3.5%（n=6）;对各成分含量间相关性分析发现环稀醚萜苷类与黄酮类组分含量间无相关性,四种环烯醚萜苷类成分的含量之间存在着显著相关性,两种黄酮类成分的含量之间也存在相关性。该方法简便、快速、准确,重现性好,更符合中药材复杂体系的多指标质量控制发展趋势,为粗茎秦艽药材的质量控制找到了简便易行的方法。

To establish an HPLC with switching wavelength method for determination of six components（ loganic acid,swertiamarin, gentiopicroside,sweroside,isoorientin,isovitexin）in Gentiana crassicaulis. Sepax Gp-C18（150 mm × 4. 6 mm,5 μm）column was used,the mobile phase was methanol-0. 1%phosphoric acid solution with gradient elution at a flow rate of 1. 0 mL·min-1 . The col-umn temperature was set at 25℃,detection wavelength was 235 nm（0～51 min）、270 nm（51SONG Jiu-hua;YANG Wen-yu;MENG Jie;LI Yao;ZENG Yu;CHENG Tao;CHEN Xing-fu57 min）、243 nm（57～65 min）、270 nm（65～80 min）. The six components（loganic acid,swertiamarin,gentiopicroside,sweroside,isoorientin,isovitexin）content in the ranges of 0. 01000 ～10. 0000,0. 00216 ～2. 1600,0. 048 ～48. 0000,0. 0010～1. 0000,0. 00034 ～0. 3400 and 0. 00026 ～0. 2600 μg respectively,had a good linearity. The average recoveries were 98. 52% ～99. 31%,the RSD was less than 3. 5%. There were significant correlations between the content of the four iridoid glucosides,the content of the two flavone constituents had corre-lations too. There was no correlations between the flavone constituents and the iridoid glucosides constituents. Conclusion：This meth-od was simple,rapid,accurate,dependable. And may be use to control quality of Gentiana crassicaulis.