Pharmacognosy- Glycosides

Theory and importance of glycosides

1. Glycosides By Subject Teacher : Mr . Dattaprasad N. Vikhe Designation : Assistant Professor Department of Pharmacognosy Course: B Pharmacy Subject :- P’cog & Phyto - II Class : Third Year B. Pharmacy

2. Organic natural compounds present in a lot of plants and some animals, these compounds upon hydrolysis give one or more sugars (glycone) moieties and non sugar (aglycone) moietyd Glycoside=sugar group+ nonsugar group (glycone) (aglycone or genin) Glycone and aglycone are linked by glycosidic linkage defination

3. Glycosidic linkage is formed between – OH group of sugar and –H group of non sugar moieties OR other sugar with loss of water molecule glycoside bond glycoside=glycone{---o---}aglycon

4. Sugar in glycosides is mostly beta-D glucose but other sugars like galactose, mannose, rhamnose, digitoxose can present. Glycosides can be alpha or beta but plants contains only beta glycosides therapeutic effect of glycosides is only due to aglycon part only and suger moiety facilitate absorption of glycoside, transportation of aglycone to site of action . so glycosides are hydrolysed to give aglycon part for action

5. Glycosides have therapeutic effect in human and animals they are used in traditional and modern medicines as cardio tonic purgative A nalgesic anti- rheumatic demulcent many other uses.

6. Classification 1-classification based on type of aglycone in glycoside Anthraquinone or anthracene glycoside Sterols or cardiac glycoside Saponin glycoside Cyanogenic glycoside Isothiocynate glycoside Coumarins and furanocoumarins Aldehyde glycoside phenol glycoside Steroidal Miscellaneous glycosides

7. 2-classification based on type of sugar (glycone) part glucose- glucoside Rhamnose – rhamnoside Digitoxose- digitoxoside Glucose and rhamnose- glucorhamnoside Rhamnose and glucose- rhamnoglucoside

8. 3-classification based on type of linkage between glycone and aglycone Linkage between –OH group of glycone and –H group of RADICALS LIKE –CH,-OH,-SH,-NH of aglycone. glycoside is named by perfix like C-glycoside,-sugar linked to carbon atom of aglycone N-glycoside,- sugar linked to nitrogen atom of aglycone O-glycoside,- sugar linked to oxygen atom of aglycone S- glycoside- sugar linked to sulfur atom of aglycone 4-classification based on there use Cathartics, cardio tonics, analgesics, anti-rheumatics, anti- ulcer e tc

9. Physical & chemical properties Colorless, solid, amorphous, nonvolatile (flavonoid- yellow, anthraquinone-red or orange.) Give positive reaction with Molisch's and Fehling's solution test (after hydrolysis). They are water soluble compounds, insoluble in organic solvents Most of them have bitter taste (except: populin, glycyrrhizin, stevioside )

10. Odorless except saponin (glycyrrhizin). when a glycosides has a lot of sugars its solubility in water decrease. Glycosides hydrolyzed by using mineral acids and temperature or by using enzymes

11. General extraction method of glycosides Sta-otto method The drug containing glycoside is finely powdered and subjected to successive extraction in a soxhlet apparatus with alcohol or suitable solvent. first take drug containing glycoside, finely powdered that, and it is extracted with alcohol or water by using soxhlet apparatus. After extraction, collect the extract and treat with lead acetate to precipitate tannins

12. after that filter it and to the filtrate pass H2S gas, to precipitated excess of lead acetate and convert it to ppt of lead sulphide as this is toxic. Now the extract again filter. The filtrate is subjected to fractional crystallization, distillation or chromatography gives pure component And molecular structure of component is determined by the spectrophotometer, Ultra Red assays, Infra red , NMR and mass spectroscopy etc.

13. Saponins glycosides They contain aglycon part sapogenin. harmful safogenins are called sapotoxins. This aglycone part have foaming action when shaked with water and yields colloidal solution. They are considerd as haemotoxic because they causes haemolysis of erythrocytes, hence some of them used as fish poisons Saponins are categorized in to two groups 1-steroidal saponins(tetracyclic triterpenoid saponins) 2-pentacyclic triterpenoid saponins

14. Chemical tests for saponin glycosides Haemolysis tes t-a drop of blood on slide was mixed with few drops of aq. Saponin solution,RBCs becomes ruptured in presence of saponins Foam test- to 1gm of drug add 10-20 ml of water, shake for few min, formation of froth which persist for 60-120 seconds

15. Liquorice roots

16. Liquorice roots marathi- jestha madhu Synonyms Glycyrrhiza, mulethi, yasti, Biological source- It consist dried ,unpeeled roots and stolons of Glycyrrhiza glabra linn Family – leguminosae It should contain not less than 3% of glycyrrhinic acid.

17. Geographical source Commercially cultivated in Spain, Sicily and England Varieties of glycyrrhiza glabra G.Glabra var. typica ( spanish liquorice ) G.Glabra var. glandulifera ( russian liquorice ) G.Glabra var. violacea ( persian liquorice )

18. Cultivation and collection Propogation of plant is done by using pieces of stolons. each piece should have 2-3 buds of aerial shoot. Plant grow well in deep sandy soil with manures or fertilizers. pieces of stolon planted in march at 2*3 distance. Crop is kept free from weeds. Roots harvested after 3-5 years of planting,they are obtained from plants by dugging befor fruiting. Drug is washed with water. Large pices are cut in small one and dried under sun and then in shade

19. Macroscopic charecters Colour- unpeeled drug yellowish brown or dark brown externally and yellowish internally Odour - faint and characteristic Taste - sweet size - length 20 to 50 cm, diameter 2cm Shape- unpeeled pieces are straight Fracture - fibrous in bark and splintery ( small, thin, sharp piece of wood) in wood Extra feature- unpeeled pieces show presence of small buds and longitudinal wrinkles

20. Microscopic characteristics

21. Cork cells- polyhedral tubular brownish colour thick, lignified, partially lignified fibers in phloem and xylem Starch and cal oxalate crystals present in parenchyma Stolon contains pith while root do not contain pith Pith , or medulla, is a tissue in the stems of vascular plants. Pith is composed of soft, spongy parenchyma cells, which store and transport nutrients throughout the plant

22. Chemical constituents Main constituent is glycyrrhizin( glycyrrhizic acid) glycoside on hydrolysis it yields glycyrrhetinic acid(triterpenoide) other constituents are sucrose, glycyramarin(bitter principle), resins, fat, asparagin it contain flavonoids like liquiritin and isoliquiritin

23. Chemical test Thick section of drug or powder+ 80% sulphuric acid=yellow colour

24. Uses Expectorent, demulcent (agent that forms a soothing film over a mucous membrane, relieving minor pain and inflammation) , falvouring agent, anti- inflammatory, rheumatide arthritis , due to flavonoid content with antigastric ulcer effect it is used in peptic ulcer, antispasmodic it is ingredient of liquorice compound powder which is potentiate the effect of senna

25. Adulterants and substitutes Manchurian liquorice ( glycyrrhiza uralensis), it is free from sugar but contains glycyrrhizin- pale chocklet brown colour Russian liquorice ( G glabra var glandulifera), it have purplish colour

26. DIOSCOREA

27. DIOSCOREA SYNONYM- yam, rheumatism root B.S- it consist of dried tubers of plant Dioscorea deltoidea,D.composita, and other species of dioscorea. family- dioscoreaceae

28. Geographical source- North western Himalaya,Punjab to Nepal, In China up to altitude of 1000 to 3000 meter. it is cultivated in Jammu and Kashmir

29. Cultivation and collection -Cultivated from tubers with crown of about 70-80 gm in weight. -first sown in nursery bed. -after 30-40 day tubers sprout.( germinate ) -after 2-3 month of growth transplanted in field at distance of 30*60cm. -they required support for optimum growth as veins are weak and tender. -Manures and organic fertilizers are supplied. -Irrigation done every 10 days -fresh tubers are harvested by deep ploughing after two years of crop. -Dried and they loose 50% weight after drying.

30. Macroscopic charecterstics Colour- slightly brown Odour- odourless Taste-bitter Extra features- it is climber, rhizomes are soft, horizontally arranged and very close to soil, drug covered with scattered roots.

31. Microscopic characteristics- Epidermis is absent in TS. cork consist few layers. and few layers of cortical parenchyma tissue. stele ( primary vascular tissue in the stem or root of a vascular p lant, consisting of the xylem and Phl o e m t o g e ther wit h supporting tissues, such as pith.) is major part of drug consisting collateral fibro-vasculer bundles. endodermis and pericyle are indistinguishable.

32. CHEMICAL CONSTITUENTS 75% non edible starch as it is bitter . chief constituent is diosgenin { sugar-free ( aglycone ) . It is hydrolytic product of saponin- dioscin } other Glycosides- smilagenin, epismilagenin, yammogenin

33. Uses Diosgenin is steroidal sapogenin so used as precursor for synthesis of corticosteroids, sex hormones, oral contraceptive. diosgenin used in treatment of rheumatoid arthritis

34. Isolation of diosgenin dried tubers are powdered and dioscin is hydrolysed by mineral acids to obtain diosgenin, then it is extracted by non polar solvents like benzene or solvent ether

35. Allied species Dioscorea flouribunda- india, central america - D.villosa - U S A - D , d e l t oi d ea w all v a r . s i kk i me n s i s p r ain Himalayas, Nepal, Sikkim Cheilocostus speciosus or crêpe ginger is alternate source of diosgenin (1.5%)

36. DIGI T ALIS

37. SYNONYMS Digitalis leaves , foxglove leaves B S it consist dried leaves of Digitalis Purpurea family Scrophulariaceae Should contain not more than 5% moisture, dried below 60 C

38. GEOGRAPHICAL SOURCE it is cultivated in England, Eu r op e , USA , India

39. CULTIVATION AND COLLECTION It is propagated by seeds. it requires calcarious, acidic, sandy soil for growth. seeds are very small in size i.e 100 seeds weigh 40 to 70 mg. seeds are mixed with fine sand and sown in nursery beds in march/april. young seedlings are transplanted in sep/november. crop is manured and kept free from weeds. In first year plant bears rosette leaves and in second year sessile leaves. Leaves collected in second year by handpicking when 2/3 rd of flowers are fully developed.

40. leaves immediately dried in vacuum driers. dried leaves containing not more than 5% moisture are packed in airtight containers with dehydrating agents. Presence of moisture deteriorates glycosides. If dried above 60 C potency lost due to chemical degradation.

41. MACROSCOPIC CHARECTERS Colour - dark greyish green odour - slight taste- bitter size - 10 to 40 cm long and 4 to 20 cm wide shape- ovate- lanceolate(shape like lance head or tapering to a point at each end) to broadly ovate( like egg) Extra feature Leaves are slightly pubescent on both surfaces, pinnate venation,(one main vein extending from the base to the tip of the leaf and smaller veins branching off the main vein.) generally leaves are broken and crumbled.( break in small fragments)

42. Microscopic characters Dorsiventral leaf.(two surfaces differing from each other in appearance and structure,) covering and glandular trichomes on boath surfaces. covering trichomes are uniseriate,( arranged in single series) 3 to 4 cells long, having collapsed cells, acute apex and warty( hard rough lump growing on) cutical. glandular trichomes are short, unicellular stalk and bicellular or unicellular head. it have anomocytic stomata (irregular celled)

43. Starch grains are present in endodermis. collenchyma present at upper and lower epidermis and pericyclic ( is a cylinder of parenchyma or sclerenchyma cells that lies just inside the endodermis and is the outer most part of the stele of plants.) part .

44. Chemical constituents 0.2 to0.45% cardenolides . purpurea glycoside A and B, glucogitaloxin. Othe r, odoroside H, gitaloxin, verodoxin, glu c o v e r od o x i n Saponin glycosides digi t oni n , gitonin

45. Hydrolysis products of purpurea glycosides Purpurea glycoside A Purpurea glycoside B hydrolysis hydrolysis digitoxigenin + 3 digitoxose gitoxigenin +3 digitoxose digitoxigenin gitoxigenin

46. Uses Congestive heart failure,( heart is unable to pump sufficiently to maintain blood flow to meet the needs of the body ) artrial fibrillation( common type of abnormal heartbeat. The heart rhythm is fast and irregular in this condition. ) , artrial flutter, ( abnormal heart rhythm that occurs in the atria of the heart . When it first occurs, it is usually associated with a fast heart rate or tachycardia (beats over 100 per minute)) supraventricular tachyca rdia

47. Ad u l t e r a n ts Verbascum Thapsus ( mullelin leaves)- large wooly branched candelabra(large branched candlestick or holder for several candles) trichomes. Primula Vulgaris (comfrey leaves)-multicellular trichomes forming hook at top. Symphytum Officinale ( primrose leaves)-uniseriate covering trichomes 8 to 9 cell long.

48. Anthraqiunone glycosides Anthraquinone is derivative of anthracene.( C 14 H 10 , consisting of three fused benzene rings) Natural anthraquinone derivative have laxative effects These glycosides contains anthraquinone, anthrone, anthranol, dianthranol, oxanthrone, dianthrone etc as aglycone part. parent molecule for all these aglycones is anthraquinone. In reduced form anthraquinone is present as anthranol or anthrone wich are isomeric to each other.

49. oxanthrone is intermediate substance from anthraquinone to anthranol. Bimeric form of anthrone is dianthrone. in fresh drug these aglycones are present in reduced form witch are biologically more active but during drying and storage they hydrolyzed , oxidized. Sugar part can be glucose, rhamnose, arabinose, primeverose etc.

50. anthraquinone a n t h r one o x a n t h r one

51. Chemical tests 1 borntrager,s test To 1 gm of drug add 5-10 ml of dil HCl, boil on water bath for 10 min and filter. Filtrate then extracted with CCl4/ benzene and add equal amount of ammonia solution to filtrate and shake. Ammonical layer becomes pink or red due to presence of anthraquinone .

52. 2 .modified borntrager,s test to 1 gm drug add 5 ml dil HCl + 5 ml ferric chloride( 5%w/v). Boil on water bath for 10 min, cool , filter. Filtrate then extracted with CCl4/benzene and add equal volume of ammonia solution. Pink or red colour forms due to anthraquinone . This test used for C- type anthraquinone glycoside

53. Indian senna

54. Senna Synonym senai- ki- patti, tinnevelley senna, cassia senna B.S It consist of dried leaflets of Cassia Angustifolia known as indian senna or Cassia Senna vahl family Leguminosae it should contain note less than 1 % of sennoside A and B on dried basis

55. GEOGRAPHICAL SOURCE inndian senna cultivated in t i nn e v ell e y , madu r a i , r amn a thp u r am di s tr i cs in Tamilnadu , kaddapa dist in Anthrapradesh , Kutch in Gujarat and Rajasthan

56. cultivation collection and preparation sowing is done by seed broadcasting method . for earlier germination seed surface is triturated with sand . seeds are sown thinly. it required red loamy or coarse gravelly soil( high proportion of small stones). First sowing done in feb- march. Second sowing done in oct- november. It required semi irrigation or light irrigation.. Once flowers are grown cutting given to flower stalk for further branching to occur. leaves are harvested after 2-3 months of planting.

57. First plucking done when leaflets fully grown, second plucking after one month of first plucking. Last plucking done after 4-6 weeks of second. Plant is uprooted after third plucking. Leaflets are dried in shade for 7-10 days. leaves are tossed( move from side to side or back and forth) to separate pods. Then packed in to large bundles under hydraulic pressure. Pressing produce transverse line on Indian senna witch are absent on Alexandrian senna leaves.

58. Macroscopic characteristics colour - yellowish green odour - slight taste- mucilaginous, bitter and charecterstic size - 7-8 mm width, 25-60 mm length shape- lanceolate, apex acute with spine at top.trichomes present on both surfaces.

59. Microscopic characterstics

60. it is isobilateral leaf.( divided into symmetrical halves ) trichomes present on both surfaces. Trichome,s are unicellular, conical, thick walled warty, slightly curved at base .palisade tissue present on upper and lower surface, they contain cluster crystals of cal oxalate . Pricyclic fibers present towards upper epidermis and above xylem

61. Chemical constituents sennoside A , sennoside B . they have rhein( cassic acid) dianthrone as aglycone. other anthraquinone sennoside C,D, rhein , kaempferol, aloe- emodin, isorhamnetin, mucilase , resin, myricyl alcohol, salicylic acid, crysophagic acid, Cal. oxalate. tinnevelley glucoside

62. USES p u r g a t i v e MOA- anthraquinone glycosides absorbed first in intestinal tract, then aglycone is separated and excreted in colon. it causes irritation and stimulation of colon, so movement( peristalsis) of colon increases, peristalsis also reduce water absorption hence soft and bulky faeces are formed. due to resin and emodin content griping occurs hence drug is given with carminatives.

63. ADULTERANTS AND SUBSTITUTES Dog senna (c. obovata )- shape of leaves is obovate(ovate with the narrower end at the base.) with tapering apex. Papillose( nipple-like structure) cells in lower epidermis Palthe senna- anthraquinone glycosides absent.leaflets have long hairs and when boiled with chloral hydrate solution crimson colour occurs. bombay senna, mecca senna, arabian senna etc have leaflets brownish green colour and more elongated and narrower in shape.

64. Alexandrian senna synonym f oli a se n na e al e x andr i n a , egy p t i an se n na , c ass i a senna B.S it consist dried leaflets of Cassia Acutifolia delile family Leguminosa

65. Macroscopic and microscopic charecters are same like indian senna. colour is pale greyish green. Base is more symmetrical, more pubescent. Chemical constituents and adulterants are same like Indian senna

66. Aloe

67. Alo e s Synonym Aloe, Hindi-musabbar, kumari. Marathi- korphad BS It is dried juice of leaves of Aloe Barbadensis miller known as curacao aloe or ALoe Perryi baker known as socotrine aloes or Aloe Ferox miller , and hybrids of this species Aloe Africana miller and Aloe Spicatya baker known as cape aloes family Liliaceae

68. Geographical source it is indigenous to estern and southern Africa. Cultivated in Caribben Island, Eu r ope, India

69. Cultivation and collection Propagation is done from root suckers. Root suckers planted in rows about 50 cm apart. Plant grow in dry climate and poor grade soil. Roots do not penetrate much in soil. Manure is provided. Leaves are cut first time after second year of planting and drug is obtained from leaves for 12 years. After 12 years plant is uprooted and new crop is taken. during collection cut is given at base of leaves so juice located in parenchymatous cells of pericycle exudes out.

70. Preparation of aloe 1 barbados or curacao aloes it is obtained by giving cut on leaves of aloe barbadensis. because of spines on leaves it is put in to kerosene tins immediately after cutting . Then kept in tilted position on V- shaped wooden troughs to drain out juice. juice is boil in copper pans for evaporation to obtain thick juice. Then it is poured it to metal containers , where it hardens.

71. 2 cape aloes Obtained from aloe ferox and its hybrid species. Leaves are cut transversely and kept in circular manner in basin shaped depression lined with goat skin or canvas. They are kept in this position for 5-6 hours till all juice exudes out and collected in goat skin. juice is boiled in iron kettle with continuous stirring with wooden paddle. Once juice is thick it is poured in to wooden cases where it solidify.

72. 3 socotrin aloes It is obtained from aloe perryi in east Africa. juice is collected in goat skin and allowed to become semisolid in nature. It is exported in past like consistency. 4 zanzibar aloes it is verity of socotrine aloe. Juice is placed in skin of small carnivorous animals, where it solidify. Then packed in wooden boxes. It is also called monkey skin aloe, although skin is not of monkey.

73. Description 1 curacao aloes Odour -strong odour resembling to iodoform. Taste is bitter. Colour is brownish black, opaque mass. Fracture -uneven fractured surface 2 cape aloes colour- dark brown or greenish brown to olive brown mass. Taste – nauseating and bitter. Odour - sour but distinct. Fracture - glassy

74. 3 socotrine aloes colour - brownish yellow, opaque mass with pasty consistency. Taste- extremely bitter, nauseous. Odour –unpleasant 4 zanziber aloes colour- liver brown. Odour - characteristic but not disagreeable. taste bitter. Fracture – dull, waxy, smooth, even

75. Microscopic charecterstics- TS shows outermost cuticle followed by epidermis, palisade tissue, mucilaginous parenchyma mesophyll. mesophyll contains vascular bundle covered with pericycle fibers layer. Inside pericycle ,aloetic cells are located witch contains aloe gel ( viscous yellow). Calcium oxalate crystals present in parenchyma. leaves are sessile

76. Microscopic chrecters of aloe powder It is useful for identification of powdered aloes. it is studied in lacto phenol reagent to gradually solubilize particles so crystals can clearly seen curacao aloes fragments consist of large number of very small needle or slender prisms cape aloes transparent, brown, angular or irregular fragments. socotrine aloes fragments consist of large prisms in group or dispersed form. zanzibar aloes irregular lumps in with modular masses are embedded.

77. Chemical constituents anthraquinone glycosides. main active constituent is Aloin. Aloin is also called as barbaloin barbaloin is C-glycoside Al o in

78. Oth e r isobarbaloin, beta barbaloin, aloe-emodin, resin, aloetic acid, homo n a t alo i n , aloesone, saponins, mucopolysccharides, glucosamines, hexuronic acid. aloe resin contains aloesin( purgative action)

79. Barbaloin( aloin) aloesin

80. Chemical teasts general chemical tests 1 gm aloe is powder boiled with 10 ml water, filtered. Filtrate used for bromine test and schoenteten,s reaction. bromine test- filtrate+ freshly prepared bromin solution= pale yellow ppt of tetrabromalin schoenteten,s test- filtrate+ borax shake well until borax dissolves.few drop of this soluition added in test tube filled with water= green fluorescence appears.

81. Special test these test are to distinguish between varieties of aloe 1 nitrous acid test- aqueous solution of aloe + sodium nitrate crystals + acetic acid curacao aloes -sharp pink to carmine colour cape aloes- faint pink colour socotrine and zanzibar aloes- very less change in colour

82. 2-Nitric acid test- nitric acid applied to drug or its aqueous solution curacao aloes- deep brownish red colour cape aloe- brownish colour changes to green socotrine aloes- pale brownish- yellow colour zanzibar aloes- yellowish brown colour

83. 3 kupraloin test( klunge,s isobarbaloin test)- Dil. aqueous solution of aloe+ drop of copper sulphat+ sodium chloride + excess 90% alcohol Curacao aloes- wine red colour persist for 4 hours Cape aloes- faint colour rapidly changes to yellow Socotrine aloes -no colour zanziber aloes- no colour

84. Uses purgative. Stronger purgative than all anthraquinon glycosides. To prevent gripping action carminative can be given. it is ingredient of compound tincture of benzoin( friar,s balsam) aloe gel is used for topical application and many cosmetic uses.

85. Aloe gel- it is obtained from inner parenchyma cells of leaf. Slightly viscous and clear liquid. It should not be c o nt amin at ed with aloe juice

86. Adulterants and substitutes Natal aloes- it contains natalion, homonatalion, resin, it is weak purgative Mocha aloes- brittle, black and glossy with strong odour Aloe can be adulterated with black catechu . Alcoholic extract of aloe gives deep brown colour while black catechu gives black colour.

87. ge n tian

88. Ge n tian Synonym - Gentiana, radix gentianae B.S It is dried partially fermented rhizome and root of yellow gentian i.e Gentiana Lutea Family Ge n tiana c eae

89. Macroscopic characteristics colour - rhizomes are yellowish-brown Odour -peculiar( different) Taste - sweet taste followed by intense bitter Fracture - short and smooth in dried drug but tough and flexible in moist drug

90. Chemical constituents Bitter glycoside -gentiopicrin(gentiopicroside) O t he r s - Amarogentin, ( bitter) ama r o sw er i n , gentioside, Gentinin Gentisin( yellow colour flavonoid) Gentisic acid, gentianose(trisaccharide) Gentibiose(disaccharide) sucrose

91. Chemical test under UV light gentian extract shows light blue fluorescence

92. Uses Bitter tonic