1. How to isolate alkaloid

2. Extraction 6 kg of dry weight Extracted with methanol (CH 3 OH) several times Exhaustive maceration technique Methanol was evaporated using rotary evaporator 600 g of methanolic extract produced and applied for the next fractionation process

3. Air Dried Plant Material Extraction with MeOH Dissolved in 0.5 M HCl Washed with CH 2 Cl 2 CH 2 Cl 2 Basified to pH 9 HCl pH9 Extraction with CH 2 Cl 2 Basified to pH 11 pH11 Extraction with CH 2 Cl 2 Acidified with HCl to pH 2 Reduce N-oxides using zinc powder Zn Basified to pH 9 Extraction with CH 2 Cl 2 Basified to pH 11 Crude Alkaloid FractionationHClpH2

4. Crude Alkaloids Fractions Fractions (mg) MeOH fraction (g) Dried plant Weigh (g) EDCBA pH11pH9pH11pH9HCl 197.7357.6211.1984.22815.5628.46161.6 +ve ++ve-ve Dragendorff reagent test Fraction A: no alkaloids present Alkaloids are presents in all other fractions Fraction B is the alkaloids rich fraction Fraction B applied for chromatographic process