Piperine can be isolated in good yield from ground black pepper (2% to 5%), which is made up of 5% to 9% alkaloids/amides that also include piperidine, piperettine and piperanine. Pungency of pepper is attributed to piperine and piperanine. Though extracted from pepper, it is manufactured synthetically for commercial uses.

A. Method I

The common procedure for the isolation of piperine involves extraction of the ground black pepper using ethanol (95%).

1. Extract 15 g of powdered pepper with 150 ml of 95% ethanol on a Soxhlet extractor for 2 h.
2. Concentrate the extract to 10-15 ml by simple distillation.
3. Treat the concentrated extract with 10 ml of 10% Potassium hydroxide in 95% ethanol (for removal of the resin fraction of the drug).
4. Heat the resulting solution and add water drop wise. A yellow precipitate is formed. Add water drops until no more precipitate is formed.
5. Allow the mixture to stand overnight and collect the solid by vacuum filtration and recrystallize from acetone.

B. Method II

1. Take 10 g of powdered pepper in a 100 ml round-bottomed flask, add 50 ml of dichloromethane and boil the mixture under reflux for 20 min.
2. Cool the flask, filter through a Buchner funnel to remove the powdered material. Wash the residue with 5 ml dichloromethane. Set aside a few drops for identification by TLC.
3. Transfer the filtrate to a rotary vacuum evaporator and remove the excess solvent until a dark brown oil is left.
4. Cool the oily liquid in an ice bath and add 10 ml cold ether. Stir well and remove the remains of solvents by gentle heating on a sand bath.
5. Add further 10 ml ether and set aside in an ice bath for 15 min with occasional stirring.
6. The precipitate of piperine formed is separated by vacuum filtration and washed with ice cold ether (2 × 5 ml).
7. For recrystallization transfer the residue on the filter into a test tube, dissolve in a hot 3:1 mixture of acetone and hexane.
8. Set aside for 15 min at room temperature and then for 30 min in an ice bath. Vacuum filter the formed crystals of piperine and record the melting point.