蒙药铁杆蒿的定性和定量分析

Qualitative and Quantitative Analysis of Mongolian Medicine Artemisa Sacrorum Ledeb

关键词:  铁杆蒿  香豆素  薄层色谱法  高效液相色谱法

Key words:  Artemisa sacrorum Ledeb  coumarin  TLC  HPLC

摘要:

目的 建立铁杆蒿的TLC鉴别及HPLC定量分析方法。方法 采用TLC和HPLC对采集于内蒙古通辽市5个不同地区的铁杆蒿进行定性和定量分析。TLC条件：对照品为8-羟基-6，7-二甲氧基香豆素和7-羟基-6-甲氧基香豆素，吸附剂为硅胶G，展开剂为二氯甲烷-丙酮（7：1），于紫外灯下显色观察；对照品为6，8-二甲氧基香豆素-7-O-β-D-葡萄糖苷，吸附剂为硅胶G，展开剂为二氯甲烷-甲醇（5：1），于紫外灯下显色观察。HPLC条件：色谱柱为Topsil C₁₈（250 mm×4.6 mm，5 μm），流动相为水（A）-乙腈（B），梯度洗脱，检测波长为263 nm，柱温为30℃。结果 对照品8-羟基-6，7-二甲氧基香豆素、7-羟基-6-甲氧基香豆素和6，8-二甲氧基香豆素-7-O-β-D-葡萄糖苷与不同样品具有Rf值相同的特征斑点，重复性好，易于鉴别；在选用的HPLC条件下，所检测化合物（1~7）表现出良好的线性关系（r ≥ 0.999 3），其平均回收率分别为98.5，95.0，97.0，98.3，96.0，98.9，99.1，RSD均<2.0%。结论 建立了一种简便、稳定和可靠的铁杆蒿定性定量分析方法，为综合评价该药材质量提供依据。

Abstract:

OBJECTIVE To establish the TLC identification and HPLC quantitative analysis method for Artemisa sacrorum Ledeb. METHODS TLC and HPLC were used for qualitative and quantitative analysis of Artemisa sacrorum Ledeb collected from five different regions in Tongliao, Inner Mongolia. TLC conditions were as follow:The reference substances were 8-hydroxy-6,7-dimethoxycoumarin and 7-hydroxy-6-methoxycoumarin, the adsorbent with silica gel G, the developing agent with dichloromethane-acetone(7:1) and the chromogenic reagent with ultraviolet lamp; The reference substance was 6,8-dimethoxycoumarin-7-O-β-D-glucoside, the adsorbent with silica gel G, the developing agent with dichloromethane-methanol(5:1) and the chromogenic reagent with ultraviolet lamp. HPLC conditions were as follow:The separation was performed on Topsil C₁₈(250 mm×4.6 mm, 5 μm); The mobile phase was comprised of water(A)-acetonitrile(B) with a gradient elution. The detection wavelength was 263 nm with column temperature at 30℃. RESULTS The spots in TLC of reference substances(8-hydroxy-6,7-dimethoxycoumarin, 7-hydroxy-6-methoxycoumarin and 6,8-dimethoxycoumarin-7-O-β-D-glucoside) and different samples had the same Rf value, with good repeatability and easy to be identified. Under the HPLC conditions adopted in this study, all calibration curves compound 1-7 exhibited good linearity(r ≥ 0.999 3). The recoveries of the method were 98.5, 95.0, 97.0, 98.3, 96.0, 98.9, 99.1, respectively. RSD were <2.0%. CONCLUSION A simple, stable and reliable method for qualitative and quantitative analysis method of Artemisa sacrorum Ledeb, which provides a basis for comprehensive evaluation of the quality of Artemisa sacrorum Ledeb.