RP-HPLC法测定桦褐孔菌中紫萁酮含量

Determination of Osmundancetone in Inonotus obliquus by RP-HPLC

建立应用反相高效液相色谱(reversed-phase high performance liquid chromatography,RP-HPLC)检测桦褐孔菌中紫萁酮含量的方法。分别采用不同的溶剂和方法对桦褐孔菌进行提取,应用HPLC对提取物中的紫萁酮进行含量测定,并进行方法学考察。色谱条件为:RP-C18色谱柱(150 mm×4.6 mm,5μm);柱温为25℃;流动相为甲醇-0.5%醋酸水(20∶80),流速0.5 mL/min;进样量5μL;检测波长320 nm。紫萁酮含量在0.5μg/mL^5μg/mL范围内与峰面积线性关系良好(r=0.999 9),该定量方法精密度、稳定性和重复性良好,其RSD分别为0.22%、0.77%和0.59%,方法的平均回收率为100.05%。该研究建立RP-HPLC测定桦褐孔菌中紫萁酮含量的方法,该方法前期处理简便,测定结果准确,精密度、稳定性、灵敏度高和重复性良好,可用于桦褐孔菌及其它天然产物提取物中紫萁酮的含量测定,为桦褐孔菌资源开发及质量评价提供一定的参考依据。

To establish a reversed-phase high performance liquid chromatography quantitative method of the determination of osmundancetone in Inonotus obliquus.Different solvents and methods were used to extract Inonotus obliquus.RP-HPLC was applied to determine the content of osmundancetone in the extract of Inonotus obliquus,and the methodology investigation was conducted.The chromatographic conditions were:RP-C18 chromatographic column(150 mm×4.6 mm,5μm),the column temperature was 25℃,the injection volume was 5μL,the mobile phase was methanol-0.5%acetic acid water(20∶80),and the flow rate was 0.5 mL/min,the detection wavelength was 320 nm.Osmundancetone showed a good linear relationship with peak area in the range of 0.5μg/mL-5μg/mL(r=0.9999).The precision,stability and repeatability of the quantitative method were better,and its RSD were 0.22%,0.77%and 0.59%,respectively.The average recovery rate was 100.05%.In this study,the RP-HPLC quantitative method and pre-treatment process of osmundancetone in Inonotus obliquus were simple and effective.The determination results were accurate,the HPLC method was with good precision and stability,high sensitivity and great repeatability.This method could be used to determine the content of osmundancetone in Inonotus obliquus and other natural product.The study could provide some reference for the development and quality evaluation of inosporium obliquus.