基于染色法和HPLC法测定龙葵不同部位中多酚含量

Determination of polyphenols in different parts of Solanum nigrum L.by staining and HPLC method

目的采用染色法测定龙葵不同部位的总酚酸含量;采用高效液相色谱(HPLC)法同时测定龙葵不同部位5个酚酸类化合物[绿原酸、咖啡酸、没食子酸、(E)-阿魏酸和原儿茶酸)]及1个黄酮化合物(芦丁)的含量。方法采用三氯化铁-铁氰化钾染色法测定龙葵不同部位的总酚酸含量;采用HPLC梯度洗脱,分段波长的方法同时测定6个多酚类成分的含量。结果染色法方法学结果显示总酚酸(没食子酸)在其线性范围内与测定值线性关系良好(r=0.9985),加样回收率为103.70%,RSD为2.7%,不同部位总酚酸以熟果中最高[(7040±195)μg·g^(-1),0.70%],其次为叶[(4430±120)μg·g^(-1),0.44%]、青果[(2720±97)μg·g^(-1),0.27%],茎中最低[(810±9)μg·g^(-1),0.08%]。HPLC方法学结果显示各成分在各自线性范围内与峰面积线性关系良好(r≥0.9993),加样回收率在95.58%~108.41%,RSD在0.34%~4.5%。结论染色法和HPLC法准确、稳定、可靠,可为龙葵药材的质量控制及物质基础研究提供依据。

Objective To determine the content of total phenolic acid in different parts of Solanum nigrum L.by staining,and to determine the contents of 5 phenolic acids[chlorogenic acid,caffeic acid,gallic acid,(E)-ferulic acid,and protocatechuic acid]and 1 flavonoid(rutin)in different parts of Solanum nigrum L.by HPLC.Methods The content of total phenolic acid in different parts of Solanum nigrum L.was determined by ferric chloride-potassium ferricyanide staining.The content of 6 polyphenols was determined by HPLC gradient elution with segmented wavelength.Results Methodology of staining showed that the linear range of the total phenolic acid(gallic acid)was good(r=0.9985),the recovery was 103.70%,and RSD was 2.7%.The highest total phenolic acid was found in the mature fruits[(7040±195)μg·g^(-1),0.70%],followed by the leaf[(4430±120)μg·g^(-1),0.44%],green fruits[(2720±97)μg·g^(-1),0.27%],and the stem[(810±9)μg·g^(-1),0.08%].The content of total phenolic acid in the mature fruits was the highest.Methodology investigation of HPLC showed good linearity(r≥0.9993).The recoveries were 95.58%~108.41%with RSD of 0.34%~4.5%.Conclusion Staining and HPLC are accurate,stable and reliable,which provides the scientific basis for the quality control and material basis research of Solanum nigrum L..