ISOLATION IDENTIFICATION of alkaloids atropine and quinine.pptx

PHYTOCHEMISTRY AND PHARMACOGNOSY II, UNIT III, ISOLATION IDENTIFICATION AND ANALYSIS OF ALKALOIDS

1. SUB – PHYTOCHEMISTRY AND PHARMACOGNOSY II UNIT III ISOLATION IDENTIFICATION AND ANALYSIS OF alkaloids Prepared by : Snehal Uttam Kashid ( Asso . professor) Department of Pharmacognosy , SPMs college of pharmacy, akluj

2. Atropine occurance Atropine is a tropane alkaloid Biological source - Atropa belladonna , Datura stramonium and Hyoscyamus niger Family – Solanaceae Chemical constituents – atropine, hyoscine , hyoscyamine , belladonin , apoatropine , norhyoscyamine . Atropine is levoratatory isomer of hyoscyamine

3. Extraction and Isolation Take weighed quantity of coarse powder and moisten with sodium carbonate solution. Extract the blended mixture in petroleum ether. Filter the petroleum ether extract Extract the filtrate with aqueous acetic acid ( alkaloids extracted in aqueous layer) Extract the aqueous fraction with solvent ether and separate both fraction. Discard solvent ether fraction

4. Extraction and Isolation Aqueous (Acidic fraction) made alkaline with sodium carbonate solution to obtain precipitates of tropane alkaloids. Filter the precipitate and dry to obtain residue. Dissolve the residue in diethyl ether . Filter and concentrate the filtrate.

5. Extraction and Isolation Atropine crystals will be separated out. Filter the crystals and dissolve in alcohol containing sodium hydroxide solution ( Hyocyamine is converted to atropine) Recrystallize the atropine sulphate from acetone. Separate the crystals of atropine

6. Chraterization of atropine Thin-layer chromatography (TLC) Sample preparation – Dissolved 1mg of Atropine in 1ml of Chloroform Stationary phase - Silica gel –G Standard sample - Atropine Detecting agent – Drangendroffs reagent to produce yellow orange Color spots Mobile phase – Toluene - Ethyl acetate – Diethyl amine (70:20:10) RF Value – Compare with standard Atropine (0.70)

7. Thin-layer chromatography (TLC)

8. Identification Chemical test – Vitalin – morin test – Take small quantity of the solid atropine and add 2 drops of Con.nitric acid in an evaporating dish evaporated to dryness on water bath. Then dissolve the residue in 1ml of acetone. Add few drops of freshly prepared alcoholic potassium hydroxide solution. Violet colouration takes place due to tropane nucleus

9. Uses Atropine is a prescription medicine used to treat the symptoms of low heart rate ( bradycardia ) reduce salivation and bronchial secretions before surgery antidote for overdose of cholinergic drugs or mushroom poisoning . Antispasmodic Mydriatic

10. QUININE It is a quinoline alkaloid of cinchona bark. Biological sources : It consists of dried inner bark of Cinchona calisaya, C.succirubra , C.officinalis , C.ledgeriana Family – Rubiaceae . Quinine and quinidine are stereo-isomers . Quinine is levorotatory and quinidine is dextrorotatory The other important alkaloids of this drug are quinidine , cinchonine , cinchonidine , cinchonamine , etc.

11. ISOLATION The dry powder bark material is first well mixed with about 30% of its weight of calcium hydroxide or calcium oxide Sufficient quantity of sodium hydroxide solution to make a paste. It is allowed to stand for few hours. The mass is then transferred to a Soxhlet apparatus and extraction is carried out with benzene

12. ISOLATION Subsequently the benzene extract is shaken with successive portions of 5% sulphuric acid. The aqueous acid extract is adjusted the pH 6.5 with dilute sodium hydroxide, cool. Crystals of neutral quinine sulphate are formed. These crystals are freed from cinchonine and cinchonidine by repeated recrystallization from hot water

13. ISOLATION Colouring matter is removed by activated charcoal. Quinine sulphate crystals are dissolved in dilute sulphuric acid and made alkaline with ammonia. Initially amorphous quinine is formed , which becomes crystalline. Finally washed to remove sodium and ammonium salts and dried to 45- 55 o C

14. Charaterization A. Thin-layer chromatography (TLC) Method Sample preparation – Dissolved about 1mg of Quinine or Cinchona alkaloid in 1ml of methanol Stationary phase - Silica gel –G Mobile phase – Chloroform – Diethylamine (9:1) Standard sample – quinine sulphate Detecting agent – Dragendroffs reagent RF Value – Quinine – 0.17, Quinidine -0.26

15. Charaterization B. Identification tests Drug + little glacial acetic acid heat in dry test tube purple vapors are produced cinchona present A greenish resinous mass obtained by treating an aqueous solution of quinine sulphate with chlorin or bromine and then adding ammonia in excess. This is an identity-test for quinine. Aqueous extract + silver nitrate gives white ppt produced which is soluble in nitric acid quinidine present.

16. Uses Quinine is antimalarial Quinidine is a cardiac depressant Quinidine used in cardiac arrhythmias. Quinidine is valuable in preventing atrial fibrillation Cinchona is bitter stomachic Anti-pyretic

17. Bibliography Kokate , C.K., Purohit , A.P. and Gohkale , S.B. (2019)  Pharmacognosy . In Terpenoids, 57th Edition, Nirali Prakashan , Pune. Pharmacognosy And Phytochemistry By Vinod Rangari . 2009 2 nd edition, Career Publications.