CARBOHYDRATES

PH 105 PHARMACOGNOSY-I

Carbohydrates are polyhydroxy aldehydes or ketones, or substances that yield such compounds on hydrolysis

These are made up of C, H and O and are the

first complex organic compounds formed in the plants as a result of photosynthesis.

These

are

the

constituents

of

various

metabolites as glycosides and required precursor for biosynthesis of all other

metabolites as well as basis of all organic

compounds of living world.

Biological importance

they provide energy through oxidation they supply carbon for the synthesis of cell components

they serve as a form of stored chemical

energy they form part of the structures of some cells and tissues

Classification
Carbohydrates Simple Sugar (Saccharide) Polysaccharide

Monosaccharid e

Disaccharide

Trisaccharide

Tetrasaccharid e

Simple Sugar
Monosaccharide

Monosacchrides general formula

are

characterized presence

by of

Cn(H2O)n,

carbonyl group and (n-1) hydroxyl group.

The number of carbon atoms most often five or six. These are simplest sugar molecules and hence cannot be hydrolyzed into simpler form.

Properties

Crystalline substances Soluble in water Practically insoluble in organic solvents like

ether, chloroform and absolute alcohols.

Optically active and exist in more than one isomeric form.

Sub-classified on the basis of no. of carbon atoms present. a. Bioses 2 carbon atom, not occur free in nature. a. Trioses 3 carbon atom e.g. glyceraldehydes b. Tetroses 4 carbon atom e.g. threose, erythrose c. Pentoses 5 carbon atom e.g. xylose, arabinose, rhamnose, ribose and ribulose. d. Hexoses 6 carbon atom e.g. glucose, fructose, mannoseand galactose.

Monosaccharaides
Trioses: Tetrose:

D-glyceraldehyde

Pentose:

Hexose:

D-Ribose

Dglucose

On the basis of presence of carbonyl group: 1. Aldoses- (containing aldehydic group) e.g. glucose, arabinose and galactose

2. Ketoses- (containing ketone group) e.g.

fructose.

Carbohydrates are found in plants in more than one isomeric form i.e. D or L Most of the natural monosachharides belongs to D series (except L-rhamnose, Larabinose and L-fucose). Monosachharides can exist in both cyclic structures i.e. either pyrano (six membered ring) or furano (five membered ring) depending on nature of bridge (1-4 or1-5).

Aldohexoses e.g. glucose

generally occurs

forms in

pyranose pyranose

whereas ketohexoses forms furanose ring,

configuration

while

fructose

occurs

in

furanose configuration

The carbon atoms in ring are of sp3 hybridization and may not be in the planner form; hence it adopts various conformations

like chair, boat, half chair etc.

Preffered pyranose

conformation is always

for

aldohexo-

chair

conformation

(most stable) which has minimal interaction and lowest energy.

Disaccharide Carbohydrate which upon hydrolysis yield two molecules of monosaccharide

Sucrose is commonly obtained from sugar cane and hence known as cane sugar. It yields glucose and fructose on hydrolysis.

Lactose is obtained from milk and known as

milk sugar, it yields glucose and galactose on hydrolysis.

Maltose is obtained by hydrolysis of starch during germination of grains, it yields two molecules of glucose on hydrolysis.

Lactulose is a synthetic disaccharide, acts

as osmotic laxative, yields galactose and fructose on hydrolysis.

On the basis of type of glycosidic linkage present the disaccharides are of two types: 1. Non reducing disaccharides (linkage

involving reducing function of both sugar)

e.g. sucrose 2. Reducing disaccharides (linkage involving reducing function of only one sugar) e.g. Lactose, maltose, cellobiose etc.

Trisaccharide Compound liberate three molecules of monosaccharide on hydrolysis

Raffino se
Hydrolysis

Hydrolysis

Gentiano se

Glucose + Fructose + galactose (Beet and sugarcane) Glucose + Glucose + Fructose (Gentian root)

Tetrasaccharide Stachyose yield four molecules of monosaccharide

Polysacchrides (Glycans)

These are high molecular weight polymers of larger number of monosaccharide molecules.

The sugar molecules are attached to each

other by glycosidic linkage between the hemi-acetal hydroxyl group on C-1 of one sugar and any of the hydroxyl group on other sugar molecules.

These serve as skeletal material (cellulose) or reserve food material (starch, glycogen and inulin).

These are non-sugar carbohydrates; do not

have sweet taste, either insoluble in water (cellulose) (starch). or forms colloidal solution

Glycans i.

are

further

sub-divided

in

two

categories: Homoglycans (homogenous

polysaccharides):

formed

by

the

condensation of larger number of same sugar molecules, e.g. starch, cellulose, glycogen etc. ii. Heteroglycans (heterogeneous

polysaccharides) e.g. gums, mucilages and

Starch from

is

main

reserve It

energy

source, of two

universal constituent of plant and derived glucose. composed

components:
a. Amylose (20-30%) is a water soluble, linear chain molecule, composed of 250- 300 -D glucose having 1-4 linkage. It forms blue color with iodine.

b. Amylopectin (70-90%), it is insoluble in cold-water, branched chain molecule, composed of about 1000 -D glucose with

1-4 and 1-6 linkage. It forms red to violet

colour with iodine. Pharmaceutical products like dextrins and cyclo-dextrins are derivatives of starch.

Cellulose is the most universal biological polymer. It is fibrous substance of cell wall and responsible for structural rigidity of plants in combination with lignin. Cellulose is a linear polymer made up of (1-4) linked D-glucose units, its molecular weight ranges from 50,000 25,00,000. Cotton fibers are the purest form of cellulose (98%), whereas oxidized cellulose and methylcellulose are cellulose derivatives used in pharmacy.

Gums are complex heterogeneous, branched and uronic acid containing polysaccharide macromolecules. These are translucent, amorphous substances, exudates of plants and produced as the result of trauma. Gums are insoluble in organic solvents but most of them are soluble in water and forms colloidal viscous solutions.

These

are

optically

active

and

dilute

solutions (<1%) precipitates upon addition of ethanol or lead sub acetate. Gums on

hydrolysis

yields

sugars

(arabinose,

galactose, glucose, mannose and xylose) with sugar acids (glucoronic acid and galacto-uronic acid).

Mucilages are normal cell constituents of high molecular weight compounds, composed mainly of sulphuric acid esters of sugar. These neither dissolves in water to form clear colloidal solutions but swells nor precipitates by addition of alcohol. Mucilages on hydrolysis yields sugar (galactose and arabinose) and sugar acids (uronic acids). Seaweed agar and carrageenan contains mucilage composed of salts of sulphate esters of polysaccharides

Pectins are defined as the group of polymers made-up of partially methylated 1-4 linked galacturonic acid residues associated with arabinan and galactan units. These are localized in middle lamella of vegetable cell wall but for industrial purpose it is obtained from inner

portion of rind of citrus fruit or from apple. It

absorbs water and swells up because of its hydrophilic property forming stiff jelly. Hence, it

Chemical Test for Carbohydrates

Mono-saccharides are the building blocks of carbohydrates. Di, oligo and polysaccharides on hydrolysis in presence of mineral acid yield monosaccharide units. Monosaccharides are soluble in water and practically insoluble in organic solvents like chloroform, ether and in absolute alcohols. These are optically active compounds and respond to various color reactions and identification tests.

i. Charring test ii. Molish test iii. Iodine test iv. Barford test v. Seliwanoffs test vi. Fehling solution test vii. Benedict test viii. Tollens test ix. Bials test x. Osazone test

1. Charring test: Carbohydrates on heating in testube or in presence of Conc. H2SO4, produces charring with smell like burning sugar. 2. Molish test: Aqueous solution of drug/carbohydrate mixed with few drops of Molish reagent (alpha naphthol) and Conc. H2SO4 was added from sidewall of testube. Formation of purple coloured ring at junction indicates presence of

3. Iodine test: It is specific for polysacchrides. Few drops of Iodine solution was added to aqueous solution of drug/polysaccharide.

Formation of blue colour, which disappears

on heating and reappears on cooling, indicates the presence of starch.

4. Barford test: This test is used to distinguish between monosacchride and disacchrides. Two ml of Barford reagent (Cupric acetate,

acetic acid and water) was added to 1 ml

aqueous minutes solution of drug and boil. of Formation of brick red precipitate in 5 indicates presence monosacchride while in 7 minutes indicates

disaccharide.

5. Seliwanoffs test: This test is used for identification of keto-hexoses or to distinguish between ketoses and aldoses. To 1 ml aqueous solution of drug, 5 ml of Seliwanoffs reagent (resorcinol in 6M HCl) was added and boiled. Formation of cherry red colour in presence of ketose (Fructose) due to formation of hydroxyl methyl furfural, which condensed with resorcinol to produce cherry red colour.

6. Fehling solution test: It is generally used for reducing sugars and composed of two solutions, which are mixed in situ. Fehling solution A composed of 0.5% of copper sulphate whereas Fehling solution B composed of Sodium Potassium Tartarate.

Equal volumes of Fehling A and Fehling B

solutions were mixed (1 ml each) and 2 ml of aqueous solution of drug was added followed

Formation

of

reddish

brown

coloured

precipitate due to formation of Cuprous oxide indicates presence of reducing sugar.

Di-,

oligo

and

poly-sacchrides

having

reducing sugars can be tested by first boiling in dilute acid solution followed by neutralization with ammonia. This neutralized aqueous is used for testing.

7. Benedicts test: It is used for reducing sugars and composed of mainly Copper sulphate and sodium hydroxide. To the 4 ml

of aqueous solution of drug, 1 ml of

Benedicts solution was added and heated almost to boiling. Formation of green, yellow, orange, red or brown colour in order of increasing concentration of simple sugar

in the test solution, due to formation of

8. Tollens test: Tollens reagent (Silver Nitrate, NaOH and Ammonia) is Ammonical Silver Nitrate (diaminesilver (I) complex), an oxidizing agent, which is itself reduced to silver metal in a clean glass reaction vessel and forms a "silver mirror", when raects with aldehydes to form carboxylic acids. Add few drops of freshly prepared Tollens reagent to 2 ml of aqueous solution of drug in clean testube and heat gently. Formation of black mirror on the sidewall of testube indicates the presence of

9. Bials test: It is used to distinguish between pentoses and hexoses. Pentoses reacts with Bials reagent (Orcinol in Conc. HCl and

traces of FeCl3 as catalyst) to form furfural,

which condenses with orcinol to produce blue-green product. Aqueous solution of drug (2 ml) was mixed with 4 ml of Bials reagent and heat to boiling, it produces blue-

green colour in presence of pentose sugar.

10. Osazone test: The osazone test was developed by Emil Fischer to identify aldose sugars differing in configuration only at the alpha-carbon. These sugars react with 2, 4dinitro-phenyl hydrazine effecting only alpha-carbon with formation of pink-red coloured bis-phenylhydrazone, known as an osazone. Application of the osazone reaction to D-glucose and D-mannose demonstrates that these compounds differ

Chemical test for Starch: It is soluble in hot water, gives positive test for Molish reagent and some specific tests like Jelly test and

Lugols iodine test.

Jelly test: To 0.5 gm of starch in a testube add 5 ml of distilled water and boil on water bath. Formation of translucent jelly indicates

presence of starch.

Lugol's iodine test: It is also known as iodine KI reagent and composed of aqueous Iodine solution in presence of KI. Few drops of iodine KI reagent was added to the aqueous solution of starch, which produces deep blue to bluish black colour due to presence of amylase. The colour developed disappears on warming and reappears on cooling. Starch amylopectin, disacchrides and cellulose do not produce

AGAR

Synonyms

Agar-agar Gelose Japan-agar

Chinese-isinglass
Bengal isinglass Ceylon isinglass Layor carang Vegetable gelatin.

Biological Source

Agar is the dried hydrophilic colloidal polysaccharide complex obtained as the dried gelatinous substance from Gelidium

amansii or various species of red algae

Family: Gelidaceae

Preparation

It is an usual practice in Japan where the red-algae is cultivated by placing poles or bamboos spread in the ocean which will

serve as a support and shall augment the

growth of algae on them.

During the months of May and October the poles are removed and the algae are

carefully stripped off from them.

The fresh seaweed thus collected is washed thoroughly extracted in in water and subsequently hot digestors containing

solution of dilute acid (1 portion of algae to

60 portions of diluted acid).

The mucilagenous extract is filtered through linen while hot and collected in large wooden troughs to cool down to ambient

temperature so as to form solid gel.

The gel is mechanically cut into bars and passed through a wire netting to form strips. The moisture from the strips is removed by

successive freezing and thawing* and finally

sun dried and stored as thin agar strips.

Description

Colour : Yellowish white or Yellowish grey Odour : Odourless Taste : Bland and mucilaginous Shape : It is available in different shapes, such as: bands, strips, flakes, sheets and coarse powder

Size :
cms

Bands: width = 4cm; Length = 40 to 50

Sheets: Width = 10-15cm; Length = 45

It is insoluble in cold water in organic solvents. It readily dissolves in hot solutions and it

forms a translucent solid mass which

characteristic is very useful in microbiology for carrying out the Standard Plate Count.

Chemical Constituents

Agar can be separated into two major fractions

(a) Agarose-a neutral gelling fraction

(b)

Agaropectina

sulphated

non-gelling

fraction. The agarose is responsible for the gelstrength of agar and consists of (+) galactose and 3,6-anhydro-()-galactose

moieties

Agaropectin is responsible for the viscosity of agar solutions and comprises of sulphonated polysaccharide wherein both

uronic acid and galactose moieties are

partially esterified with sulphuric acid.

Chemical Tests
1. It gives a pink colouration with Ruthenium Red solution.

2. A 1.5-2.0% (w/v) solution of agar when

boiled and cooled produces a stiff-jelly.

3. Prepare a 0.5%(w/v) solution of agar and add to 5 ml of it 0.5 ml of HCl, boil gently for 30 minutes and divide into two equal

portions:
(a) To one portion add BaCl2 solution and observe a slight whitish precipitate due to the formation of BaS04 (distinction from Tragacanth),

(b) To the other portion add dilute KOH solution for neutralization, add 2 ml of Fehlings solution and heat on a water bath.

The appearance of a brick red precipitate

confirms the presence of galactose.

Substituents/Adulterant

Substituents/Adulterants for agar.

Gelatin

and

isinglass are usually used as substituents

Uses

used in making photographic emulsions, employed as a bulk laxative, used in preparing gels in cosmetics. It is widely used as thickening agent in confectionaries and dairy products. used in the production of ointments and medicinal encapsulations. In microbiology, it is employed in the preparation of bacteriological culture media. Used in the dyeing and printing of fabrics and textiles. Used as dental impression mould base.

PECTIN

Pectin is a group of polysaccharides found in nature in the primary cell walls of all seed bearing plants and are invariably located in the middle lamella.

It has been observed that these specific polysaccharides actually function in

combination
substance.

with

both

cellulose

and

hamicellulose as an intercellular cementing

Pectin is naturally found in a number of plants namely: lemon peel, orange peel, apple pomace, carrots, sunflower-heads,

guava, mangoes and papaya.

Biological Source

Pectin

is

the

purified

admixture

of

polysaccharides, obtained by carrying out the hydrolysis in an acidic medium of the

inner part of the rind of citrus peels Citrus

aurantium

family Rutaceae

Preparation

lemon/orange rind or apple pomace; besides the attempt to prepare either low methoxy group or high methoxy group pectins.

Preserved or freshly obtained lemon peels are gently boiled with approximately 20 times its weight of fresh water maintained

duly at 90C for a duration of 30 minutes.

The effective pH (3.5 to 4.0) must be maintained with food grade lactic acid/citric acid/tartaric extraction. acid to achieve maximum

Once the boiling is completed the peels are mildly

squeezed to obtain the liquid portion which is then

subjected to centrifugation to result into a clear solution.

From this resulting solution both proteins and starch

contents hydrolysis. are suitably removed by enzymatic

The remaining solution is warmed to deactivate the

added enzymes. The slightly coloured solution is effectively

decolourized with activated carbon or bone charcoal

resulting solution both proteins and starch contents are suitably removed by enzymatic hydrolysis. The remaining solution is warmed to deactivate the added enzymes. The slightly coloured solution is effectively decolourized with activated carbon or bone charcoal. The pectin in its purest form is obtained by precipitation with water-miscible organic solvents (e.g., methanol, ethanol, acetone), washed with small quantities of solvent and dried in a vaccum

Description
Appearance : Coarse or fine- powder Colour : Yellowish white Odour : Practically odourless Taste : Mucilaginous taste Solubility : 1. Completely soluble in 20 parts of water forming a solution containing negatively charged and very much hydrated particles. 2. Dissolves more swiftly in water, if previously moistened with sugar syrup, alcohol, glycerol or if first mixed with 3 or more parts of sucrose.

Chemical Constituents

Pectin occurs naturally as the partial methyl ester of a (14) linked (+) polygalacturonate sequences interrupted

with (12) () rhamnose residues.

The neutral sugars that essentially form the side chains on the pectin molecules are namely: (+) galactose, () arabinose, (+) xylose, and () fructose.

Chemical Tests
1. A 10% (w/v) solution gives rise to a solid gel on cooling. 2. A transparent gel or semigel results by the interaction of 5 ml of 1% solution of pectin with 1 ml of 2% solution of KOH and subsequently setting aside the mixture at an ambient temperature for 15 minutes. The resulting gel on acidification with dilute HCl and brisk shaking yields a voluminous and gelatinous colourless

Uses

It is employed mostly as an intestinal demulscent. It is believed that the unchanged molecules of polygalacturonic

acids may exert an adsorbent action in the

internal layers of the intestine, thereby producing a protective action along with Kaolin to prevent and control diarrhoea.

As a pharmaceutical aid pectin is used frequently as an emulsifying agent and also as a gelling agent preferably in an acidic

medium.

It is employed extensively in the preparation of jellies and similar food products e.g., jams, sauces, ketchups.

Poectin in the form of pastes exerts a bacteriostatic activity and hence, is used frequently in the treatment of indolent ulcers

and deep wounds.

A combination of pectin and gelatin find its application as an encapsulating agent in various pharmaceutical formulations to afford sustained-release characteristics.