高效液相色谱法同时测定山蜡梅叶中的l-洋蜡梅碱、槲皮素和山奈酚的含量

摘要

运用高效液相色谱法,对江西道地药材山蜡梅叶中l-洋蜡梅碱、槲皮素和山奈酚3种成分进行含量测定,采用Agilent HC-C18色谱柱(250 mm×4.6 mm,5μm);流动相为甲醇-磷酸盐水溶液(10:90,v/v)至(90:10,v/v)梯度洗脱;柱温:25℃;流速1.0 mL/min;检测波长0-15 min为239 nm,16-45 min为365 nm;三个主要成分分离度良好,在测试范围内呈良好的线性关系,R2≥0.9991;精密度、重复性、稳定性良好;平均加样回收率在97.47％-98.26％之间,RSD平均值小于2.27％.利用该方法对不同采收时间的山蜡梅药材进行了生物碱和黄酮成分的定量测定,结果表明4～7月份药材中生物碱含量随月份推后不断增加,而黄酮成分在4、5月份的含量最高,这与民间选择春季采药的习惯相匹配.该方法具有准确、快速、简便的特点,为山蜡梅药材的质量控制方法研究提供了实验数据,对于保证其质量和临床疗效具有重要的意义.%A reversed-phase high-performance liquid chromatography (RP-HPLC) method was established for the simultaneous determination of three major constituents,l-calycanthine,quercetin,and kaempferol,in Chimonanthi Nitentis Folium.The RP-HPLC analysis was carried out using an Agilent HC-C18 (4.6 mm×250 mm,5 μm) column and a gradient mobile phase consisting of methanol and phosphate buffer (10:90→90:10,v/v) at a flow rate of 1.0 mL/min.The column temperature was 25 ℃ and the detection wavelength was set at 239 nm from 0 to 15 min and 365 nm from 16 to 45 min.All calibration curves showed good linearity (R2≥0.9991) within test ranges.Relative standard deviations of repeated analyses were less than 2.64％ (n =6),and the recovery of this method was 97.47％-98.26％.The contents of these three analytes were determined for the samples from different harvest times.The results showed that thel-calycanthine content in herbs increases from 99.94 μg/g to 468.0 μg/g from April to July,whereas the content of quercetin and kaempferol was higher in April and May than that in June and July,which were consistent with the Chinese traditional medicine's use of Chimonanthi Nitentis Folium in Spring.Herein,a simple,rapid and accurate analytical method is presented and successfully applied for the simultaneous quantitative analysis of alkaloids and flavones from Chimonanthus nitens Oliv.