三叶青片剂的质量标准研究

Study on quality standard of Tetrastigma hemsleyanum Tablets

目的研究三叶青片剂的质量标准,为其质量控制奠定基础。方法采用薄层色谱(TLC)法鉴别三叶青片剂中荭草苷、异荭草苷、牡荆素、牡荆素鼠李糖苷及异牡荆素;采用三乙胺显色紫外-可见分光光度(UV-Vis)法,测定三叶青片剂中总黄酮的含量;建立高效液相色谱(HPLC)法测定三叶青片剂中8个主要成分(荭草苷、异荭草苷、牡荆素、牡荆素鼠李糖苷、异牡荆素、绿原酸、新绿原酸及隐绿原酸)的含量,Welch Ultimate XB-C 18色谱柱(4.6 mm×150 mm,2.7μm),梯度洗脱,检测波长:330 nm,流速:0.4 mL·min-1。结果薄层色谱法鉴别三叶青片剂中荭草苷、异荭草苷、牡荆素、牡荆素鼠李糖苷、异牡荆素的展开剂为乙酸乙酯∶甲酸∶甲苯∶水=8∶1∶1∶1,AlCl 3显色后紫外光365 nm下检视;紫外-可见分光光度法测定总黄酮含量,以牡荆素计,在1.85~11.1μg·mL-1范围内与吸光度值呈良好的线性关系,平均加样回收率为98.43%,RSD为2.46%。建立的高效液相色谱法,在考察的浓度范围内,8个成分的浓度与峰面积呈良好的线性关系(r>0.9990),平均回收率为96.88%~99.25%,RSD为1.94%~2.47%。结论本试验建立薄层色谱法鉴别三叶青片剂中荭草苷、异荭草苷、牡荆素、牡荆素鼠李糖苷及异牡荆素,用紫外-可见分光光度法测定三叶青片剂中总黄酮含量,用高效液相色谱法测定8个主要成分含量,为三叶青片剂的质量控制提供参考。

Objective To study the quality standard of the Tetrastigma hemsleyanum Tablets(THT),and provide the foundation for its quality control.Methods Using thin-layer chromatography(TLC)to identify orientin,isoorientin,vitexin,vitexin-2-O-rhamnoside and isovitexin in THT;Triethylamine color development with UV-Vis spectrophotometry was used to determine the content of total flavonoids in the THT;High performance liquid chromatography(HPLC)was established to determine 8 major components(orientin,isoorientin,vitexin,vitexin-2-O-rhamnoside,isovitexin,chlorogenic acid,neochlorogenic acid and cryptochlorogenic acid),Welch Ultimate XB-C 18 column(4.6 mm×150 mm,2.7μm),gradient elution,detection wavelength:330 nm,flow rate:0.4 mL·min-1.Results TLC to identify orientin,isoorientin,vitexin,vitexin-2-O-rhamnoside,and isovitexin in the THT using the developing agent ethyl acetate∶formic acid∶toluene∶water=8∶1∶1∶1,after AlCl 3 color development,inspection under ultraviolet light 365 nm;UV-Vis method to determine the total flavonoids content,there is a good linear relationship with the absorbance value at the range of 1.85~11.1μg mL-1 of vitexin,and the average recovery was 98.43%with the RSD of 2.46%.The established HPLC method showed a good linear relationship between the concentration of the 8 components to their peak areas(r>0.9990)within the investigated concentration.The average recovery was 96.88%~99.25%,and the RSDs were 1.94%~2.47%.Conclusion In this experiment,the TLC method for THT was established to identify orientin,isoorientin,vitexin,vitexin-2-O-rhamnoside and isovitexin in the THT;UV-Vis was used to determine the content of total flavonoids,and HPLC Methods was used to determine the content of 8 components in the THT,which provided a reference for the quality control of it.