Identification and isolation of betulin, betulinic acid and lupeol from birch bark

Betulae cortex, Betula pendula Roth., Betulaceae , comprise triterpene substances which are confirmed to possess very important pharmacological activities such as anti-inflammatory, anticancer and antiviral. In this study, acetone-methanol extraction of triterpene substances from both, inner and external birch bark was carried out. Insolubility in water of triterpenes betulin, betulinic acid, lupeol and oleanolic acid, is used for precipitation of triterpene substances from methanolic extracts. Qualitative analysis of triterpenes in precipitated raw extracts was performed by method of thin layer chromatography, applying system for development, benzene – ethyl acetate – formic acid (36:12:5). Separated spots of betulin (Rf 0.62), betulinic acid (Rf 0.59) and lupeol (Rf 0.75) were colored in shadows of violet color by reagents anisaldehyde sulphuric acid and did not show fluorescency (UV lamp 254 and 366nm). Dry column chromatography and preparative thin layer chromatography were used to isolate triterpene substances from row triterpene mixture. The study include all obtained IC specters and interpretations on the basis of which can be concluded that triterpene substances, betulin, betulin acid and lupeol isolated from external birch bark give identical characteristic signals and absorbance as those referent. Betulinic acid – IR (KBr) (ν, cm -1 ): 3484 (OH); 2870 (CH 2 ); 1686 (C=O); 1432 (OH); 1362 (CH 2 =CH-CH 3 ); 1156 (C-O), Betulin – IR (KBr) (ν, cm -1 ): 3448 (OH); 2868 (CH 2 i CH 3 ); 1638 (C=C); 1432 (OH); 1388 (CH 2 =CH-CH 3 ), Lupeol – IR (KBr) (ν, cm -1 ): 3308 (OH); 2872 (CH 2 ); 1630 (C=C); 1468 (CH); 1380 (CH 3 ) „Umbrella„; 920 (CH alkenes). Method of dry column chromatography has resulted as simple, efficient, repeatable and economical for laboratory conditions. References: [1] Krasutsky, P.A. (2006) Nat. Prod. Rep. 23:919–942. [2] Dzubak, P. et al. (2006) Nat. Prod. Rep. 23:394–411.