OBJECTIVE:To establish the quality standard of total triterpenoids in Diospyros kaki leaves. METHODS:In situ pretreatment-TLC was used for the qualitative identification of total triterpenoids(oleanolic acid and ursolic acid). With the reference of ursolic acid,spectrophotometry was used to determine the content of total triterpenoids. HPLC was used to determine the contents of oleanolic acid and ursolic acid;the column was COSMOSIL5 C18 with mobile phase of methanol -0.2% phosphoric acid (82 ∶18,V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 210 nm,the column temperature was 25 ℃,and the injection volume was 10 μl. RESULTS:TLC of oleanolic acid and ursolic acid showed clear spots and good separation. The linear range of total triterpenoids was 7.52-45.12 μg/ml. The linear range was 18.4-184 μg/ml for oleanolic acid(r=0.999 9)and 18.8-188 μg/ml for ursolic acid(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 2%;recoveries were 98.83%-102.11%(RSD=1.21%,n=6)and 99.00%-103.80%(RSD=1.82%,n=6),respectively. CONCLUSIONS:The standard can be used for the quality control of total triterpenoids in D. kaki leaves.
关键词
柿叶总三萜齐墩果酸熊果酸薄层色谱法分光光度法高效液相色谱法
Keywords
Total triterpenoids in Diospyros kaki leavesOleanolic acidUrsolic acidTLCSpectrophotometryHPLC