藏药麻黄中麻黄碱含量测定及物种分类上的应用

Determination of ephedrine content in Ephedra and it’s application on species identification

目的:建立麻黄中麻黄碱含量测定方法并用麻黄碱含量对藏药麻黄物种进行分类,评价特有麻黄植物藏麻黄和广泛分布的山岭麻黄的药用价值,以及麻黄碱含量随海拔变化分布特征。方法:从青藏高原采集5种麻黄17个居群样品,采用HPLC法建立了麻黄碱、伪麻黄碱含量的测定方法,以盐酸麻黄碱和盐酸伪麻黄碱作为对照品,运用XTERRA^® Phenyl柱(4.6 mm×250 mm,5μm)为分离柱,甲醇-0.092%磷酸水溶液(0.04%三乙胺、0.02%二正丁胺)1.5:98.5为流动相,检测波长210 nm,流速0.5 mL/min,柱温30℃。结果:建立的HPLC分析体系线性范围良好(r>0.9997),精密度、稳定性、重复性RSD均小于2%,加样回收率分别为盐酸麻黄碱96.65%,伪麻黄碱76.26%。在17个麻黄居群样品中中麻黄的麻黄碱含量最高,山岭麻黄中最低,藏麻黄和草麻黄中麻黄碱含量中等。麻黄碱和伪麻黄碱含量在种间无显著差异,且与海拔高度无显著相关性,但却得到了与分子系统发育进化关系相似的结果。结论:麻黄碱和伪麻黄碱为麻黄物种分类提供参考依据。海拔不是影响麻黄碱含量的主要因素。

The content of ephedrine in Ephedra and it's application on Ephedra species for identification,and the characters of changing in ephedrine content with altitude were studied by using HPLC method with 17 samples collected from different altitudes and the medicinal value of E.saxatilis and E.gerardiana was evaluated.HPLC method was as follows:XTERRA^® Phenyl column(4.6mm×250mm,5μm)used as separation column,1.5:98.5(v/v)of methanol/water consisting of 0.092%phosphoric acid(0.04% triethylamine and 0.02%dibutylamine)as mobile phase at a flow rate of 0.5 mL/min,30℃ of column temperature and 210nm of UV detector.The established HPLC analysis system has a good linear range(r>0.9997)with precision,stability and repeatability with less than 2%.The recovery rates of ephedrine hydrochloride and pseudoephedrine hydrochloride were 96.65%and 76.26%,respectively.The ephedrine content in 17 populations E.saxatilis has the highest ephedrine content whereas E.gerardiana has the lowest content of ephedrine.There was no significant difference in the content of ephedrine and pseudoephedrine between Ephedra species and also no significant correlation with altitude.However,the results were similar to the molecular phylogenetic evolution.Ephedrine and pseudoephedrine can be used in Ephedra species identification,but the altitude is not the main factor affecting ephedrine content.