PHARMACOGNOSY 3 RD CLASS 2 ND SEMESTER Lab

PHARMACOGNOSY 3 RD CLASS, 2 ND SEMESTER Lab. 7 INDOLE ALKALOIDS

INTRODUCTION Ø Indole alkaloids are a class of alkaloids containing a structural moiety of indole. Ø Indole is an aromatic heterocyclic organic compound. It has a bicyclic structure

INTRODUCTION HARMALA ALKALOIDS: - Ø Peganum harmala of the family Nitrariaceae Ø It is a woody, perennial, succulent shrub, native to arid regions. The leaves are bright green, finely divided and about 1 cm long. Both the roots and seeds contain significant quantities of beta-carbolines (indole) alkaloids. Which are absent from the rest of the plant.

Ø Botanical name: Peganum harmala Ø Family name: Nitrariaceae Ø Part used: seed Ø Local name: ﺍﻟﺤﺮﻣﻞ

Ø The round seed capsules measure about 1– 1. 5 cm in diameter, have three chambers and carry more than 50 seeds.

TRADITIONAL / MEDICINAL USES: Ø This plant has a long history of traditional uses including as dye ((turkey red)) is a color that was widely used to dye cotton in the 18 th and 19 th century and as incense from ancient times. Ø Peganum harmala seeds were known to possess hypothermic and essentially hallucinogenic properties since it is MAO (monoamine oxidase) inhibitor agent.

Ø Many studies were conducted to antibacterial, antifungal and antiviral effects of Peganum harmala seeds. Ø In Moroccan it is used as traditional medicine. Ø Seed powder is used sometimes for skin and subcutaneous tumors.

THE ACTIVE ALKALOIDS OF HARMAL SEEDS A. Harmane B. Harmine C. Harmaline D. Harmalol E. Tetrahydroharmine F. Vasicine & Vasicinone (quinazoline derivatives)

ISOLATION OF THE HARMALA ALKALOIDS Aim: - To isolation the harmala alkaloids Equipments and reagents: 1) Large beaker 9) 2%HCL 2) Reflux instrument 10) Ammonium hydroxide solution 3) Water bath 11) Chloroform 4) Large evaporating dish 12) Methanol 5) Separatory funnel 6) Litmus paper 7) Petroleum ether 8) 90 % ethanol

Procedure: - 1) Take 50 gm. of harmala seeds. 2) Macerate with 500 ml of petroleum ether over night. 3) Filter off the maceration solvent of the seeds. 4) And then reflux with 500 ml of 90 %ethanol for one hour. 5) Evaporate the resultant filtrate on the steam bath to syrup liquid. 6) Add to the concentrated filtrate 5 ml of 2%HCL. 7) Remove all the remaining alcohol by evaporating in large evaporating dish on the steam bath. 8) The acidic extract is then cooled, filtered and placed in a separatory funnel where it is made alkaline by adding ammonium hydroxide solution (according to litmus paper).

9) The alkaline solution is then extracted three times with 100 ml of chloroform. 10) The chloroform solution is then concentrated to give the product crude alkaloid (fraction A). 11) Add 10 ml methanol to fraction A, then filter, the result is two fractions, fraction B (filtrate), fraction C (precipitate).

Results: Fraction A contains the crude alkaloids. Fraction B contains (vasicine and deoxyvasicine alkaloids) Fraction C contains (harmine and harmaline).

Discussion: 1) Maceration process is for defatting process. 2) Use of reflux condenser since the plant is not affected by heat. 3) Use of methanol is to separate some of harmala alkaloids, since some are soluble in methanol and others are not.

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