GLYCOSIDES Classification Of GLYCOSIDES Chemical Tests Glycosides

Introduction: Number of medicinal plants containing organic constituents in conjugation with a sugar moiety
It can be l or 2 such compounds are called as glycosides.
They exert therapeutically significant effects on humans and animals Traditionally used in modern medicine because of their cardiotonic, purgative, analgesic, anti-arrhythmic, and demulcent action.
Definition:-Glycosides are defined as organic compounds from plants and animal sources which on enzymatic hydrolysis give one or more sugar moieties along with a non-sugar moiety.
Sugar moiety is called glycon and non-sugar moiety is called a-glycon.
Classification Of GLYCOSIDES�
BASED ON THE CHEMICAL NATURE OF NON-SUGAR MOIETY
BASED ON THE NATURE OF SUGAR MOIETY
BASED ON LINKAGE BETWEEN GLYCON AND AGLYCON PORTION.
BASED ON THE THERAPEUTIC NATURE OF GLYCOSIDE
BASED ON THE CHEMICAL NATURE OF NON SUGAR MOIETY
a) Anthraquinone glycoside anthraquinone moiety as aglycon. Ex: senna
b) Sterol or cardiac glycoside: aglycon portion is a steroid molecule. Ex: digitalis molecule
e) Saponine glycoside
e) Isothiocynate glycoside Ex: black mustard.
d) Cyanogentic glycoside Ex: white cherry bark
f) Flavonoid glycoside Ex: Ruta graveolens, citrus bio
g) Coumarin glycoside or Furano coumarine glycoside:- Ex: celery fruit cost
h) Aldehyde glycoside Ex: vanilla pods
i) Phenol glycoside Ex salcive
J) Steroidal glycoside
k) Glucosidal bitter or miscellaneous glycoside Ex salix species
Glucoside: sugar portion is glucose
Rhamnoside: sugar portion is rhamnose
Pentoside sugar portion is pentose
Fructoside sugar portion is fructose
Arabinoside sugar portion is arabinose
O-glycosides: Sugar molecule is combined with phenol or –OH group of aglycon, for example, higher plants: senna, rhubarb, Amygdaline, Salicin, cardiac glycosides, anthraquinone glycosides like sennosides etc.
Glycone –O H+HO − aglycone → Glycone −O− Aglycone + H2O
2. N-glycosides: Sugar molecule is combined with N of the –NH (amino group) of aglycon, for example, nucleosides
Glycone –OH+H N − aglycone → Glycone− N − Aglycone + H2O
3. S-glycosides: Sugar molecule is combined with the S or SH (thiol group) of aglycon, Isothiocyanate glycosides: for example, Sinigrin from black mustard
Glycone –OH+H S − aglycone → Glycone− S − Aglycone + H2O
4. C-glycosides: Sugar molecule is directly attached to the C—atom of aglycon, for example, Anthraquinone glycosides like Aloin, Barbaloin, Cascaroside and Flavone glycosides, etc. Cochineal colouring matter- carminic acid
Glycone –OH+H C − aglycone → Glycone− C − Aglycone + H2O
Chemical Tests for Anthraquinone Glycosides.
a) Borntrager’s test.
b) Modified Borntrager’s test.
Chemical Tests for Saponin Glycosides
a) Haemolysis test.
b) Foam test
Chemical Tests for Steroid & Triterpenoid Glycoside .
a) Liebermann Burchard test
b) Salkowaski test
c) Antimony trichloride test
d) Zimmermann test
Chemical Tests For Cardiac Glycoside
a) Keller-kiliani test
b) Legal test
c) Baljet test

1. GLYCOSIDES 1 Mr Nandakishor B. Deshmukh. Assistant Professor Shraddha Institute Of Pharmacy, Kondala Zambre , Washim . Subject - Pharmacognosy And Phytochemistry -I Class- B-pharm- II Sem IV

2. 2 SR NO Content 01 Introduction 02 Definition 03 Classification Of GLYCOSIDES 04 GENERAL CHARACTERISTICS 05 ISOLATION OF GLYCOSIDE 06 Chemical Tests Glycosides

3. GLYCOSIDES Introduction: Number of medicinal plants containing organic constituents in conjugation with a sugar moiety It can be l or 2 such compounds are called as glycosides. They exert therapeutically significant effects on humans and animals Traditionally used in modern medicine because of their cardiotonic, purgative, analgesic, anti-arrhythmic, and demulcent action. 3

4. GLYCOSIDES Definition:- Glycosides are defined as organic compounds from plants and animal sources which on enzymatic hydrolysis give one or more sugar moieties along with a non-sugar moiety . Sugar moiety is called glycon and non-sugar moiety is called a- glycon .   4

5. Classification Of GLYCOSIDES BA SED ON THE CHEMICAL NATURE OF NON-SUGAR MOIETY BASED ON THE NATURE OF SUGAR MOIETY BASED ON LINKAGE BETWEEN GLYCON AND AGLYCON PORTION. BASED ON THE THERAPEUTIC NATURE OF GLYCOSIDE 5

6. 1 ) BASED ON THE CHEMICAL NATURE OF NON SUGAR MOIETY a) Anthraquinone glycoside anthraquinone moiety as aglycon. Ex: senna b) Sterol or cardiac glycoside : aglycon portion is a steroid molecule. Ex: digitalis molecule e) Saponine glycoside e) Isothiocynate glycoside Ex: black mustard. d) Cyanogentic glycoside Ex: white cherry bark f) Flavonoid glycoside Ex: Ruta graveolens, citrus bio g) Coumarin glycoside or Furano coumarine glycoside:- Ex: celery fruit cost h) Aldehyde glycoside Ex: vanilla pods i ) Phenol glycoside Ex salcive J) Steroidal glycoside k) Glucosidal bitter or miscellaneous glycoside Ex salix species 6

7. 2 ) BASED ON TE NATURE OF SUGAR MOITY Glucoside: sugar portion is glucose Rhamnoside : sugar portion is rhamnose Pentoside sugar portion is pentose Fructoside sugar portion is fructose Arabinoside sugar portion is arabinose 7

8. 3) BASED ON LINKAGE BETWEEN GLYCON AND AGLYCON PORTION:- 1. O-glycosides: Sugar molecule is combined with phenol or –OH group of aglycon, for example, higher plants: senna, rhubarb, Amygdaline, Salicin, cardiac glycosides, anthraquinone glycosides like sennosides etc. Glycone –O H+H O − aglycone → Glycone −O− Aglycone + H 2 O 2. N-glycosides: Sugar molecule is combined with N of the –NH (amino group) of aglycon, for example, nucleosides Glycone – OH+H N − aglycone → Glycone − N − Aglycone + H 2 O 3. S-glycosides: Sugar molecule is combined with the S or SH (thiol group) of aglycon, Isothiocyanate glycosides: for example, Sinigrin from black mustard Glycone – OH+H S − aglycone → Glycone − S − Aglycone + H 2 O 4. C-glycosides: Sugar molecule is directly attached to the C—atom of aglycon, for example, Anthraquinone glycosides like Aloin, Barbaloin, Cascaroside and Flavone glycosides, etc. Cochineal colouring matter- carminic acid Glycone – OH+H C − aglycone → Glycone − C − Aglycone + H 2 O 8

9. 4) BASED ON THERAPEUTIC NATURE OF GLYCOSIDE:- Cardiac glycoside Ex: Digitalis laxative glycoside Ex: Senna Anti-ulcer glycoside Ex : Liquorice Bitter glycoside Ex: quassia wood 9

10. GENERAL CHARACTERISTICS 1) Glycoside contains sugar but still the physical, chemical and therapeutic activity is based on the aglycon portion. Sugar facilitates the absorption of the glycoside helping it to reach the site of action 2) Glycoside are a crystalline, amorphous substance that is soluble in water , and dilutes alcohol but is soluble in the CHC13 and ether . The aglycon moiety is insoluble in a non-polar solvent like C6H6 3) Glycosides are easily hydrolyzed by mineral acids water and enzyme They show optical activity normally they are levorotatory . 4) Glycoside can not reduce fehling’s solution until they are hydrolyzed . 5) They are believed to facilitates the growth and protection of plant. 10

11. ISOLATION OF GLYCOSIDE The drug containing glycoside is finely powdered and subjected to successive extraction in a soxhlet apparatus with alcohol or a suitable solvent. After extraction collect the extract and treat it with lead acetate to precipitate tannins after that filter it and the filtrate passes H2S gas. no lead acetate in the precipitate as lead sulphide as this is toxic. Now after the extract again filter . the extract molecular structure is determined by the spectrophotometer Ultra Red assays Infra-red, NMR mass spectroscopy etc. 11

12. Chemical Tests Glycosides Chemical Tests for Anthraquinone Glycosides. a) Borntrager’s test. b) Modified Borntrager’s test. Chemical Tests for Saponin Glycosides a) Haemolysis test. b) Foam test 12

13. Chemical Tests Glycosides Chemical Tests for Steroid & Triterpenoid Glycoside . a) Liebermann Burchard test b) Salkowaski test c) Antimony trichloride test d) Zimmermann test 13

14. Chemical Tests Glycosides Chemical Tests For Cardiac Glycoside a) Keller- kiliani test b) Legal test c) Baljet test d) 3,5-dinitro benzoic acid test 14

15. Chemical Tests Glycosides Chemical Tests for Flavonoid Glycosides a) Ammonia test b) Shinoda test c) Vanillin HCl test 15

16. Chemical Tests Glycosides Chemical Tests For Coumarin Glycosides a) FeCl 3 test b) Fluorescence test Chemical Tests for Cynophoric Glycoside a) Sodium picrate test 16

17. Chemical Tests for Anthraquinone Glycosides Borntrager’s test 1 gm of drug + 5–10 ml dil. HCl, boil on a water bath for 10 min, filter. Extract filtrate with CCl 4 / benzene. Add an equal amount of NH 3, and shake. The pink or red colour in the ammonical layer is due to anthraquinone moiety. Modified Borntrager’s test 1 gm of drug +5 ml dil. HCl + 5 ml FeCl 3 (5% w/v). Boil for 10 min on a water bath, cool and filter, extract filtrate with CCl 4 / benzene and add an equal volume of NH 3 solution→ formation of pink to red colour due to anthraquinone moiety. This is used C-type of anthraquinone glycosides. 17

18. Chemical Tests for Saponin Glycosides 1. Haemolysis test A drop of blood on slide+ mix with a few drops of aq. Saponin solution → RBC’s rupture in presence of saponins. 2. Foam test 1 gm drug + 10–20 ml H 2 O, shake for few minutes → frothing persists for 60–120 s in the presence of saponins. 18

19. Chemical Tests for Steroid & Triterpenoid Glycoside Libermann Burchard test Evaporate the alcoholic extract of the drug to dryness, extract with CHCl 3 + a few drops of acetic anhydride followed by conc. H 2 SO 4 from sides of test tube → violet to blue coloured ring at the junction of two liquids, indicating the presence of steroid moiety. Salkowaski test Evaporate the alcoholic extract of the drug to dryness, extract with CHCl 3 + conc. H 2 SO 4 from the sidewall of the test tube to extract → yellow coloured ring at the junction of two liquids, which turns red after 2 min, indicating the presence of steroid moiety. 19

20. Chemical Tests for Steroid & Triterpenoid Glycoside Antimony trichloride test Evaporate alcoholic extract of the drug to dryness, extract with CHCl 3 + saturated solution of SbCl 3 in CHCl 3 containing 20% acetic anhydride → Formation of pink colour on heating indicates the presence of steroids and triterpenoids. Zimmermann test Meta dinitrobenzene solution + alcoholic solution of drug containing alkali + heat → violet colour in the presence of keto steroid 20

21. Chemical Tests For Cardiac Glycoside Keller- kiliani test Alcoholic extract of drug + equal vol. of H 2 O + 0.5 ml of strong lead acetate solution shake, filter. Filtrate + eq. vol. CHCl 3 . Evaporate CHCl 3 extract to dryness, Dissolve the residue in 3 ml of gel. CH 3 COOH few drops of FeCl 3 solution. Transfer the resultant solution to a TT containing 2 ml conc. H 2 SO 4 → A Reddish brown layer is formed and turns bluish-green after standing due to the presence of digitoxose . Legal test Alcoholic extract of drug + equal vol. of H 2 O + 0.5 ml of strong lead acetate solution, shake, filter. Filtrate + eq. vol. CHCl 3 . Evaporate CHCl 3 extract to dryness, Dissolve the residue in 2 ml of pyridine and sodium nitroprusside 2 ml + NaOH to make alkaline → pink colour in the presence of glycosides or aglycone moiety. 21

22. Chemical Tests For Cardiac Glycoside Baljet test A thick section of a leaf of digitalis or the part of a drug containing cardiac glycoside + sodium picrate solution → yellow to orange colour in the presence of aglycones or glycosides. 3,5-dinitro benzoic acid test Alcoholic sol. Drug + drops of NaOH+ 2% sol. of 3,5-dinitro benzoic acid → pink colour indicates the presence of cardiac glycosides. 22

23. Chemical Tests for Flavonoid Glycosides a. Ammonia test Dip Filter paper in alcoholic sol. of drug + expose to NH 3 → yellow spot filter paper b.Shinoda test an alcoholic extract of drug + Mg turning + dil. HCl → red color (Flavonoids) alcoholic extract of drug + Zn turning +dil. HCl → deep red to magenta colour , presence of dihydro flavonoids. c. Vanillin HCl test Vanillin HCl + alcoholic sol. of drug → pink color due to the presence of flavonoids. 23

24. Chemical Tests For Coumarin Glycosides FeCl 3 test Conc. alcoholic extract of drug + few drops of alcoholic FeCl 3 solution → deep green colour , turns yellow on the addition of conc. HNO 3 → presence of coumarins Fluorescence test Alcoholic extract of drug + 1N NaOH (1 ml each) → blue-green fluorescence → shows the presence of coumarins. 24

25. Chemical Tests for Cynophoric Glycoside Sodium picrate test Powdered drug + moistened with H 2 O + drops of conc. H 2 SO 4 in C Flask Filter paper impregnated with sodium picrate solution followed by Na 2 CO 3 sol. trapped on the neck of the flask using cork → brick red colour due to volatile HCN in the presence of pyrophoric glycosides 25

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