Identification of phytoconstituents by Dr.U.Srinivasa, Professor and Head, Srinivas college of pharmacy, Mangalore

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Identification of phytoconstituents by Dr.U.Srinivasa, Professor and Head, Srinivas college of pharmacy, Mangalore

  1. 1. IDENTIFICATION OFPHYTOCONSTITUENTS Dr.U.Srinivasa, M.Pharm,Ph.D
  2. 2. TESTS FOR STEROIDSSALKAOWSKI TESTTo the chloroform solution in a test tube concentratedsulphuric acid was added from the sides of the test tube.A reddish brown color was observed. LIEBERMAN BURCHARDT TEST To the chloroform solution in a test tube a few drops ofacetic anhydride was added and mixed well.1ml ofconcentrated sulphuric acid was added from the sides ofthe test tube and allowed to stand. A reddish ring wasformed at the junction of two layers.
  3. 3. TESTS FOR TRITERPENES(a) SALKAOWSKI TEST A few drops of concentrated sulphuric acid were added to the chloroform solution, shaken and allowed to stand. Lower layer turned yellow.(b) LIEBERMAN BURCHARDT TEST To the chloroform solution a few drops of acetic anhydride and 1ml of concentrated sulphuric acid was added. A deep red color was produced.
  4. 4. (C)TRICHLORO ACID AND STANNIC CHLORIDE TEST To the chloroform solution a few drops of thionyl chloride and a pinch of stannic chloride were added. A range of colors green, blue, purple and finally turning to red were obtained.(D) KAHLENBERG TEST To 0.2 ml of the chloroform solution a few drops of antimony pentachloride and chloroform was added. A deep purple color was observed.
  5. 5. TESTS FOR SAPONINS FOAM TEST : Small amount of extract was shaken in a testtube with a little quantity of water, the foam producedpersisted for 10 minutes. This confirms the presence ofsaponins.TESTS FOR LACTONES(a) LEGAL’S TEST : To the extract mixture of sodium nitroprusideand pyridine was added. The mixture was treated withsodium hydroxide. It gave a deep red color. (b) BAL JETS TEST : To the various extracts treated withsodium picrate solution. Yellow to orange color wasproduced shows the presence of lactone ring.
  6. 6. TESTS FOR ALKALOIDS (A) MAYER’S TEST The various extracts were dissolved in chloroform.The chloroform was evaporated and the residue was acidified and added few drops of Mayer’s reagent (Potassium Mercuric Iodide). A creamy white precipitate was observed. (B) WAGNER’S TEST The various extracts were dissolved in chloroform. The chloroform layers were evaporated, to the residue were acidified and added few drops of Wagner’s reagent (Iodine in Potassium Iodide). Orange precipitate was observed.
  7. 7. (C) DRAGENDROFF’S TEST The various extracts were dissolved in chloroform. Thechloroform was evaporated and the residue was acidified andadded few drops of Dragendroff’s reagent (PotassiumBismuth Iodide). Orange red precipitate was observed.(D) HAGER’S TESTThe various extracts were dissolved in the chloroform. Thechloroform was evaporated and the residue was acidified andadded few drops of Hager’s reagent (Saturated Picric Acidsolution). Yellow crystalline precipitate was observed.
  8. 8. TEST FOR CARBOHYDRATES(A) MOLISCH’S TEST The extract with Molisch’s reagent mix and added concentrated sulphuric acid along the sides to form layers. A reddish violet ring at the interference shows the presence of carbohydrates.(B) FEHLING’S TEST The extract was heated with Fehling’s A and B solution it gave an orange red precipitate shows the presence of reducing sugar.
  9. 9. (C) BENEDICT’S TEST With Benedict’s reagent the carbohydrates on boiling and cooling a green reddish brown precipitate was formed which shows the presence of reducing sugar(D) BARFOEDS TEST To the extract Barfoeds reagent was added and it was boiled on a water bath, reddish precipitate was observed within 90 minutes show the presence of monosaccharide.
  10. 10. TEST FOR FLAVANOIDS:(A) FERRIC CHLORIDE TEST To a small quantity of the alcoholic solution of the extract a few drops of neutral ferric chloride solution was added. A green color was produced due to the phenolic nucleus.(B) SHINODA TEST To the alcoholic solution of the extract a few fragments of magnesium ribbon were added. To this concentrated hydrochloric acid was added drop wise. Magneta color was produced after few minutes which are the characteristic reaction of flavanoid.
  11. 11. (C) ZINC-HYDROCHLORIDE ACID REDUCTION TEST To the alcoholic solution of the extract a pinch of zinc dust was added and few drops of concentrated hydrochloric acid were added. Magneta color was produced after a few minutes.(D) LEAD ACETATE TEST To the alcoholic solution of the extract few drops of lead acetate solution (10%) was added. Yellow precipitate was observed.
  12. 12. TEST FOR TANNINS:(A) FERRIC CHLORIDE TEST 2 ml of extract was taken in a test tube and ferric chloride solution was added drop by drop. A blue black precipitate was observed.(B) GELATIN TEST To the extract few drops of 1% solution of gelatin containing 10% sodium chloride was added. A white precipitate was observed. These tests confirmed the presence of tannins.
  13. 13. TEST FOR PROTEINS:(A) BIURET TEST Reagent 40% sodium hydroxide and dilute copper sulphate solution. Protein shows blue, pink or violet color where as amino acids fails to show the color. The extract does not give this reaction. Shows the absence of protein and amino acids.(B) NINHYDRIN TEST With Ninhydrin amino acids show blue color. But proteins may give the positive tests very rarely. The extract does not give the Ninhydrin reaction.
  14. 14. TEST FOR GLYCOSIDES:(A) BAL JETS TEST To the various extracts with sodium picrate solution. Yellow to orange color was observed which shows the presence of glycoside with lactones ring.(B) KELLER-KILLANI TEST To the various extracts 1ml of glacial acetic acid and few drops of ferric chloride solution was added and then slowly concentrated sulphuric acid was added through the sides of the test tube. A reddish brown ring at the junction of liquids was observed which shows the presence of de-oxysugar.
  15. 15. TESTS FOR VOLATILE OILS1. Volatile oil is soluble in alcohol (90%)2. Put one drop of volatile oil on the filter paper, no permanent stain indicates the presence of volatile oilTESTS FOR FIXED OIL AND FATS1. Press the extract in between the two filter papers , a permanent stain indicates the presence of fixed oil2. Extract is treated with few drops of 0.5N potassium hydroxide and few drops of phenolphthalein and heat , formation of soap indicates the presence of fixed oil and fats
  16. 16. GUMS AND MUCILAGES1. Powder is treated with few drops of Ruthenium red solution , the particles acquires pink colour2. Powder when treated with water or aqueous KOH , the particles swells3. Aqueous extract with few drops of dilute Hcl and equal volumes of Fehlings solution A and B and heat, red colour is produced indicates the presence of gums
  17. 17. ESTIMATION OF PHYTOCONSTITUENTS ALKALOIDS GLYCOSIDES TANNINS VOLATILE OILS RESINS
  18. 18. GLYCOSIDESCARDIAC GLYCOSIDES :1. By colorimetric method2. By biological assay
  19. 19. THE DIFFERENCE BETWEEN A SUCCESSFUL PERSONAND OTHERS IS NOT A LACK OF STRENGTH, NOT ALACK OF KNOWLEDGE, BUT RATHER A LACK IN WILL.

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