CHEMICAL TESTS FOR GLYCOSIDES AND ALKALOIDS.pptx

Chemical tests for Glycosides & Alkaloids. Types of glycosides is classified as glycone part & aglycone part.
Test for glycosides include Borntragers test, Foam test, Libermann burchard test, Salkowaski test, Antimony trichloride test, Trichloroacetic acid test, Zimmermann test, Keller-killiani test, Legal test, Baljet test, Dinitro benzoic acid test, Fecl3 test, Fluorescence test, Sodium pictrate test, Ammonia test, Shinoda test, Vannillin test.
Types of alkaloids as True alkaloids, Proto alkaloids, Pseudo alkaloids.
Other sources of alkaloid other than Castoramine, Muscopyridine, Pyocyanine.
Chemical tests for alkaloids are Dragendroffs test, Mayers test, Hagers tst, Wagners test.

1. CHEMICAL TESTS OF GLYCOSIDE & ALKALOID Dr. Aswin Anandh . M PG SCHOLAR HOMOEOPATHIC PHARMACY

2. GLYCOSIDE Organic natural compounds present in a lot of plants and some animals,these compounds upon hydrolysis give one or more sugars (glycone) B- form andnon sugar (aglycone) or called genin.

3. CLASSIFICATION On the basis of glycone : • Glucoside: If glucose group is present in glycoside, then it is known as glucoside. • Fructoside : If fructose group is present in glycoside, then it called fructoside . • Glucuronide: If glucunoric acid is present , then it is known as glucunronide .

4. On the basis of aglycone : on the basis of presence of nature of aglycone group classes are as follows: • Sterol and triterpenoid glycosides • Anthraquinone glycosides • Cyanogenetic and cyanophoric glycosides • Saponin glycosides • Flavone glycosides • Coumarin glycosides • Steroidal or cardiac glycoside

5. CHEMICAL TEST FOR GLYCOSIDES

6. BORNTRAGER’S TEST To 1 gm of drug add 5–10 ml of dilute HCl boil on water bath for 10 min and filter. Filtrate was extracted with CCl 4 / benzene and add equal amount of ammonia solution to fil trate and shake. Formation of pink or red colour in ammoni-cal layer due to presence of anthraquinone moiety ANTHRAQUINONE GLYCOSIDE

7. FOAM TEST To 1 gm of drug add 10–20 ml of water, shake for few minutes, Formation frothing which persists for 60–120 s in presence of saponins . SAPONIN GLYCOSIDES

8. Libermann burchard test Alcoholic extract of drug was evaporated to dryness and extracted with CHCl 3 , add few drops of acetic anhydride followed by conc. H 2  SO 4  from side wall of test tube to the CHCl 3  extract.  Formation of violet to blue coloured ring at the junction of two liquid, indicate the presence of steroid moiety. Chemical Tests for Steroid and Triterpenoid Glycosides  

9. Salkowaski test Alcoholic extract of drug was evaporated to dryness and extracted with CHCl 3 , add conc. H 2  SO 4  from sidewall of test tube to the CHCl 3  extract.  Formation of yellow coloured ring at the junction of two liquid, which turns red after 2 min, indicate the presence of steroid moiety. Antimony trichloride test Alcoholic extract of drug was evaporated to dryness and extracted with CHCl 3 , add saturated solution of SbCl 3  in CHCl 3  containing 20% acetic anhydride. Formation of pink colour on heating indicates presence of steroids and triterpenoids

10. Trichloroacetic Acid test Sample + addition of saturated solution of trichloro acetic acid Forms Colored precipitate Zimmermann Test Meta dinitrobenzene solution was added to the alcoholic solution of drug containing alkali, on heating  It forms violet colour in presence of keto steroid.

11. Keller- kiliani test   To the alcoholic extract of drug equal volume of water and 0.5 ml of strong lead acetate solution was added, shaked and filtered. Filtrate was extracted with equal volume of chloroform. Chloroform extract was evaporated to dryness and residue was dissolved in 3 ml of glacial acetic acid followed by addition of few drops of FeCl 3  solution. The resultant solution was transferred to a test tube contain- ing 2 ml of conc. H 2 SO 4 .  Reddish brown layer is formed, which turns bluish green after standing due to presence of digitoxose .   CARDIAC GLYCOSIDES

12. LEGAL TEST To the alcoholic extract of drug equal volume of water and 0.5 ml of strong lead acetate solution was added, shaked and filtered. Filtrate was extracted with equal volume of chloroform and the chloroform extract was evaporated to dryness. The residue was dissolved in 2 ml of pyridine and sodium nitropruside 2 ml was added followed by addition of NaOH solution to make alkaline Formation of pink colour in presence of glycosides or aglycon moiety.

13. BALJET TEST Thick section of leaf of digitalis or the part of drug con- taining cardiac glycoside, when dipped in sodium picrate solution It forms yellow to orange colour in presence of aglycones or glycosides. 3,5-dinitro benzoic acid test To the alcoholic solution of drug few drops of NaOH followed by 2% solution of 3,5-dinitro benzoic acid was added. Formation of pink colour indicates presence of cardiac glycosides.

14. FeCl3 test To the concentrated alcoholic extract of drug few drops of alcoholic FeCl 3  solution was added. Formation of deep green colour, which turned yellow on addition of conc. HNO3, indicates presence of coumarins . Fluorescence test The alcoholic extract of drug was mixed with 1N NaOH solution (one ml each). Development of blue-green fluo-rescence indicates presence of coumarins . COUMARIN GLYCOSIDES

15. SODIUM PICTRATE TEST Powdered drug was moistened with water in a conical flask and few drops of conc. Sulphuric acid was added. Filter paper impregnated with sodium picrate solution followed by sodium carbonate solution was trapped on the neck of flask using cork. Formation of brick red colour due to volatile HCN in presence of cynophoric glycosides takes place. CYNOPHORIC GLYCOSIDE

16. Ammonia test Filter paper dipped in alcoholic solution of drug was exposed to ammonia vapor Formation of yellow spot on filter paper SHINODA TEST To the alcoholic extract of drug magnesium turning and dil. HCl was added, formation of red colour indicates the presence of flavonoids. To the alcoholic extract of drug zinc turning and dil. HCl was added Formation of deep red to magenta colour indicates the presence of dihydro flavonoids FLAVONOID GLYCOSIDE

17. Vannilin HCl Test Vanillin HCl was added to the alcoholic solution of drug, Formation of pink colour due to presence of flavonoids.

18. ALKALOIDS "Basic nitrogenous compound, contains one or more nitrogen in heterocyclic ring , system having marked p hysiological action on human and animals when use in small quantities“ .

19. CLASSIFICATION T rue alkaloids, P rotoalkaloids, P seudo alkaloids. True alkaloids and protoalkaloids are produced from amino acids, whereas pseudoalkaloids are not derived from these compounds.

20. DRAGENDROFF’S TEST Drug solution + Dragen droff's reagent(Potassium Bismuth lodide ), Formation of Orange red colour MAYER’S TEST Drug solution + few drops of Mayer's reagent (K2Hgl4) Formation of creamy-white precipitant. HAGER’S TEST Drug solution + few drops of Hagers reagent ( Saturatedaq . Solution of Picric acid Formation of crystalline yellow precipitate WAGNER’S TEST Drug solution + few drops of Wagner's reagent ( dilutelodine solution) Formulation of reddish-brown precipitate.

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