How to isolate alkaloid

Extraction

• 6 kg of dry weight

• Extracted with methanol (CH3OH) several times

• Exhaustive maceration technique

• Methanol was evaporated using rotary evaporator

• 600 g of methanolic extract produced and applied

for the next fractionation process

Air Dried Plant Material

Extraction with MeOH

Dissolved in 0.5 M HCl

Washed with CH2Cl2

CHCH22ClCl22Basified to pH 9

HClHCl

pH9pH9

Extraction with CH2Cl2

Basified to pH 11

pH11pH11

Extraction with CH2Cl2

Acidified with HCl to pH 2

Reduce N-oxides using zinc powder

ZnZn

Basified to pH 9

Extraction with CH2Cl2

Extraction with CH2Cl2

Basified to pH 11

Crude Alkaloid Fractionation

HClHClpH2pH2

Crude Alkaloids Fractions

Dried plant Weigh (g)

MeOH fraction(g)

Fractions (mg)

A B C D EHCl pH9 pH11 pH9 pH11

6161.6 628.4 2815.5 984.2 211.1 357.6 197.7Dragendorff reagent test -ve ++ve +ve +ve +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