Tannins

Definition: Organic non nitrogenous polyphenol compounds derived from Tanning

(conversion of animal hides to leather) positive goldbeater's skin test

Characters:

- Astringent water soluble compounds with high molecular weight (1000 - 5000)

- Tannins Aqueous soln are acidic to litmus paper (due to presence of free phenolic and
carboxylic groups)

- Ppt alkaloids, gelatin and heavy metals (Antidote in case of poisoning)

Y
- Show color reaction with Dichromate and iron salts.
TH
- Ppt protein making it resistant to proteolytic enzymes

- Most of tannins present in nature as glycosides

- Immature fruits contain tannins which disappear after ripening

FA

- Dicot contains Hydrolysable tannins while plants contains condensed tannins

Extraction

1) Fresh or Lyophilized drying are extracted with water + Acetone mixture

2) Filtrate is distilled to remove acetone

3) Aqueous extract is extracted with dichloromethane to remove lipids and fats

4) The remaining aqueous layer is extracted with ethyl acetate to separate (low M.wt)

Dimeric proanthocyanidins and gallotannins

5) The remaining aqueous layer contains (high M.wt) polymeric proanthocyanidins and

gallotannins

6) Further separation and fractionation done through chromatography

pg. 1
 Goldbeater's skin test:

1) Small piece of ox intestine (gold beater skin) soaked in dil HCL and washed

2) Skin soaked in test soln for few minutes, then rinsed with distill water

3) Transfer to 1% soln of Ferrous Sulphate

- Formation of brownish black color indicates presence of True Tannins

- Pseudo tannins give negative result

Tannins

Hydrolysable Condensed
Y Complex Psedo tannins
TH
Gallo tannins Ellagi tannins
A) Hydrolysable Tannins
FA

- Esters of sugars (Glucose) and phenolic acids (Gallic acid - Ellagic acid)

- Hydrolysis with Tannase enzymes alkali or with Acids to give Gallic acid or
HexaHydroxyDiphenic Acid (HHDP)

- Dry distillation gives Pyrogalol which gives Bluish black color with FeCI3

Negative with Bromine water

1) Gallotannins

- Glucose as central core which esterified by 5-6 Gallic acid

- Simple tannins present in Rhubarb, Clove and chestnut

- Molecular weight (1000-1500)

Chinese Tannins contains 5 galloyl esters while Turkish contains 4 galloyl esters

pg. 2
Hamamelis tannins in Hamamelis leaf contains:

Hamamelose (Hydroxy methyl ribose) sugar

With 2 galloyl esters

2) Ellagitannins

-Central Glucose esterified by dimer of gallic acid called HexaHydroxyDiPhenic acid (HHDP)

-Hydrolysis gives glucose and HHDP which undergoes lactonization into Ellagic acid

- Present in Pomegranate bark with molecular weight 1000 3000

Ex: Corilagin in Eucalyptus leaf

Y
TH
FA

B) Condensed Tannins

- Resist hydrolysis and doesn't contain sugars with M.wt 1000-3000

-Derivatives of Catechin, Epichatechins (isomer) Flavon-3-ol & Flavon-3,4-diol

-On treatment with Acids or enzymes converted into red water insoluble Plobaphene

- Dry distillation gives catechol which gives Greenish black color with FeCl3

Gives Ppt with bromine water

Cinnamon bark. Cinchona and green tea

Flavon-3-ol

pg. 3
C) Complex Tannins

- Consists of Hydrolysable tannin unit and condensed tannin unit connected to glucose core
through C-C bond between CI of glucose and C6 or C8 of Flavan- 3-ol

- Dry distillation gives Pyrogallol + Catechol which give bluish and greenish black color with
FeCL3

Camelliatannins in Camellia japonica

D) Pseudo tannins

Called Proto tannins or Tannins precursor

with molecular weight (200-600)

- Negative with Goldbeaters skin test

Y
TH
-Ex: Gallic acid and chlorogenic acid in Nux Vomica seed

Tannic acid
FA

Called Gallotannic acid obtained from (abnormal growth of Quercus infectoria)

- Extraction from powder with water + Alcohol + Ether and separation of aqueous layer

Witch contains Tannic acid

- Consists of Glucose and Gallic acid

- Dextrorotatory Acidic yellowish white Odourless water and alcohol soluble powder

Medicinal uses of Tannins & Tannic acid

1) Anti-diarrheal & astringent in burns, skin ulcers, gangrene and sore throat
2) Antioxidant inhibits lipids peroxidation, Anti-viral, Anti-bacterial and anti-inflammatory
3) Antidote for alkaloids and heavy metals poisoning Industrial applications includes Ink, Dye
and hide tanning
Tannins Assay
Using Poly Vinyl Poly Pyrrolidone (PUPP) which binds with tannins (Gravimetric)

pg. 4
 Terpene lactones

1) Artemisinin

Class: peroxides Sesquiterpene lactone derivative

Source: aerial parts of Artemisia annua F: Compositae

Uses: Anti-malarial drug

2) Santonin

Class: Sesquiterpene lactone derivative

Y
Source: Un-expanded flower heads of Artemisia cilla
TH
(Santonica flower) Worm seed family: Compositae

Uses: Anthelmintic for round worm with less effect on

thread worms and no effect on tape worms

FA

3) Cantharidin

Class: Monoterpene lactone drvative

Source: beetle of Lytta vesicatoria (Spanish fly) F: Meloideae

cantharides extract contains 0.6-.1 % Cantharidin

Uses: Rubefacient (Vesicant) and counter irritant in

Rheumatism and in Alopecia

- Aphrodisiac, abortifacient, but it is highly toxic (5mg) as is

excreted through kidney unchanged result in damage and
irritation for urogenital organs

pg. 5
4) Picrotoxin

Class: Mixture of two oxidized Sesquiterpene (Picrotin &

Picrotoxinin)

- Picrotoxinin is the active part

Source: Berries of Anamirta coculus Family: Menispermaceae

Uses: Antidote for Barbiturates toxicity but with narrow

therapeutic index

- Toxic for fish

Y
TH
5) Parthenolide

Class: Sesquiterpene lactone (Gerrmachanolide)

FA

Source: Tanacetum Parrchenium leaf (feverfew)

Uses: Prophylactic against Migraine (inhibit serotonin release from

platelets)

pg. 6
 Anti oxidants
Oxidation: Electron transfer from substance to another to produce free radicals

Oxidative stress: Equilibrium failure between creation and elimination of free

radicals

Oxidative stress

Result in Cell damage & LDL oxidation result in atherosclerosis CNS disorders

Antioxidant: molecule capable of slowing or prevention of oxidation of other

molecules by terminating the chain reaction

Antioxidants uses:

Anti-cancer & preservative in foods vitamins and plasticizers

Antioxidants types:

Y
a) Water soluble reacts with oxidant in cytoplasm and plasma

b) Lipid soluble protect cell membrane from lipid peroxidation ex: vitamin E
TH
c) Exogenous cannot be synthesized inside body, obtained from diet ( Vit C & E )

d) Endogenous Enzymes inside cells

FA

Antioxidants assay

ORAC (Oxygen Radical Absorbance Capacity) measure antioxidant capacity of antioxidant in

food, juice

DPPH (DiPhenyl Picryl Mydrazine) Violet free radical react with antioxidant to give yellow
color intensity assay spectrophotometrically

A) Exogenous anti-oxidants

Ascorbic acid (vit C)

- Monosaccharide maintains the reduced form of Glutathione (water soluble)

- promotes stress resistance in plants

Α Tocopherols (vit E)
- Protect cell membranes from oxidation by reacting with lipid radicals produced in lipid
peroxidation chain reaction

- Small dose is required as it stored inside body

pg. 7
 8) Endogenous anti-oxidants

1) Dismutase &Catalase & peroxi redoxin

- catalyze the break down of:

1)Super oxide into Hydrogen peroxide and oxygen

2) Hydrogen peroxide into water and oxygen

- need Cu & Zn & Fe cofactors

2) Glutathione

- Tripeptide of Glutamic-Cysteine-Glycine

- can be synthesized from diet (Endogenous)

- maintained in reduced form by Glutathione Reductase system

- Active part: Thiol group of Cysteine Y

TH
3) Melatonin

- Hormone synthesized from Tryptophan (endogenous)

- Powerful antioxidant can easily cross cell membrane and blood brain barrier
FA

(CNS)

- No Redox Cycle (Suicidal Antioxidant or Terminal Anti-Oxidants) Not reduced or

regenerated as it forms stable end products

Pro-Oxidants

Uitamin C reduce metal ions (Fe) that generate free radicals through Fenton reaction

Physical Exercise

Oxygen consumption during exercise, result in free radicals generation &

Muscular fatigue

- This strengthens the body antioxidants defense and protective against diseases associated

with oxidative stress

- Vitamin C acts as antioxidant (small dose) so increase energy output

pg. 8