How to isolate alkaloid

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

Crude Alkaloid Fractionation

Air Dried Plant Material Acidified with HCl to pH 2 Extraction with MeOH Reduce

N

-oxides using zinc powder Dissolved in 0.5 M HCl

HCl

Basified to pH 9 Washed with CH 2 Cl 2 Extraction with CH 2 Cl 2 Basified to pH 9

CH 2 Cl 2

Basified to pH 11 Extraction with CH 2 Cl 2 Basified to pH 11 Extraction with CH 2 Cl 2 Extraction with CH 2 Cl 2

Crude Alkaloids Fractions

Fractions (mg) Dried plant Weigh (g) MeOH fraction (g) A HCl B pH9 C pH11 D pH9 E pH11 6161.6

628.4

Dragendorff reagent test 2815.5

-ve 984.2

++ve 211.1

+ve 357.6

+ve 197.7

+ve • 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