HPLC法同时测定14个产地娑罗子中4种七叶皂苷

Simultaneous determination of four constituents in Aesculi Semen from fourteen different growing areas by HPLC

目的建立HPLC法同时测定14个产地娑罗子Aesculi Semen中七叶皂苷A、B、C、D的含有量。方法娑罗子甲醇提取液的分析采用Waters Xbridge C18柱(4. 6 mm×250 mm,5μm);流动相乙腈-水(含0. 2%磷酸);体积流量1. 0 mL/min;检测波长220 nm;柱温25℃。用DPS、EXCEL软件进行数据分析。结果七叶皂苷A、B、C、D分别在1. 0～10μg/mL (R^2=0. 999 4)、1. 0～10μg/mL (R^2=0. 999 5)、1. 0～10μg/mL (R^2=0. 999 6)、0. 2～2. 0μg/mL(R^2=0. 999 6)范围内线性关系良好,平均加样回收率分别为100. 01%、101. 49%、100. 73%、99. 65%,RSD分别为2. 11%、0. 68%、0. 69%、1. 54%。14个产地间娑罗子中七叶皂苷类成分含有量存在显著(P <0. 05)或极显著(P <0. 01)差异,七叶皂苷A含有量均满足2015年版《中国药典》规定标准,其中陕西、山西、河南产娑罗子中七叶皂苷类成分含有量较高,浙江产娑罗子含有量较低;七叶皂苷含有量与纬度、海拔呈正相关关系,与经度、种子纵径、种子横径呈负相关关系。结论该方法准确稳定,重复性好,可用于娑罗子的质量控制。

AIM To establish an HPLC method for the simultaneous content determination of aescin A,aescin B,aescin C,aescin D in Aesculi Semen from fourteen different growing areas. METHODS The analysis of methanol extract of Aesculi Semen was performed on a 25 ℃ Waters Xbridge C18 column(4. 6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-water(containing 0. 2% phosphoric acid) flowing at 1. 0 mL/min in a gradient elution manner,and the detection wavelength was set at 220 nm. The obtained data were analyzed by EXCEL and DPS software. RESULTS Aescin A,aescin B,aescin C,aescin D showed good linear relationships within 1. 0-10 μg/mL(R^2= 0. 999 4),1. 0-10 μg/mL(R^2= 0. 999 5),1. 0-10 μg/mL(R^2= 0. 999 6),0. 2-2. 0 μg/mL(R^2= 0. 999 6),whose average recoveries were 100. 01%,101. 49%,100. 73%,99. 65% with the RSDs of 2. 11%,0. 68%,0. 69%,1. 54%,respectively. There were significant differences of aescin contents and growth traits among Aesculi Semen from various growing areas(P < 0. 05,P < 0. 01). The aescin A contents of fourteen treatments met the specified standard of 2015 《Chinese Pharmacopoeia》. The aescin contents of Aesculi Semen from Shaanxi,Shanxi,and Henan provinces were higher than those from Zhejiang province. Moreover,there were significantly positive correlation between aescin contents and latitude,as well as height while negative correlation with longitude,vertical diameter and transverse diameter of seeds. CONCLUSION This accurate,stable and reproducible method can be used for the quality control of Aesculi Semen.