Biochem Lab Report: Carbohydrates

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The passage discusses different types of carbohydrates, reagents used to test for carbohydrates, and tests performed to identify different carbohydrates.

Arabinose, glucose, fructose, sucrose, and starch are mentioned. Arabinose, glucose, fructose, and sucrose are miscible in water while starch is immiscible in water.

Molisch's reagent, Seliwanoff's reagent, Phloroglucin, Fehling's A and B reagents, Moore's reagent, Barfoed's reagent, Tauber's Benzidine reagent, and Bial's Orcinol reagent are described along with their compositions.

Mary Angelic T.

Bihag, BSMT 2B
EXPT NO. 3 CARBOHYDRATES

1. What can you say about the solubility of carbohydrates in water?

SAMPLE STRUCTURAL FORMULA SOLUBILTIY IN


CARBOHYDRATES CLASSIFICATION WATER

Arabinose
Missible in water

C5H10O5

Missible in water
Glucose
C₆H₁₂O₆

Missible in water
Fructose C6H12O6

Missible in water
sucrose

C12H22O11

Starch
Immissible in water

(C6H10O5)n
2. Give the composition of the following Reagents

Reagents Composition

Molisch Molisch’s reagent:1 5% alpha-naphthol solution in ethyl alcohol

Seliwanoff Seliwanoff’s reagent contains (water), resorcinol, concentrated HCL


Tollen’s reagent contains concentrated HCl mixed with 0.5%
Phloroglucin
phloroglucinol

Fehling’s A Fehling’s A reagent contains aqueous copper sulfate solution.

Fehling’s B reagent contains alkaline potassium tartarate (Rochelle


Fehling's B salts)

NOT IN THE ACTIVITY

Moore's test
Moore’s reagent contains Concentrated NaOH

Barfoed's test
Barfoed’s reagent contains Copper Acetate in Glacial Acetic Acid

Tauber's Benzidine test


Tauber's Benzidine reagent contains benzidine solution.

Bial’s reagent is made by dissolving orcinol in concentrated HCL and


Bial's Orcinol Test ferric chloride solution.

3. Give the principles of the following tests:


Tests Principle

This test is based on the reaction of the alpha-naphthol with


Molisch’s test carbohydrate in the presence of sulfuric acid. The sugars react with
alpha-naphthol in an acidic environment to form purple-colored
furfural or hydroxymethylfurfural derivatives. The intensity of the
color is directly proportional to the amount of carbohydrate present in
the solution.

This test involves the formation of furfural derivatives by


Seliwanoff’s test monosaccharides with hydrochloric acid. The furfural derivatives
formed by a sugar with ketonic functional group condense with
resorcinol to form a chromogen having cherry-red color.
This test also involves the formation of furfural derivates in the
Phloroglucin test presence of concentrated HCL. The furfural derivatives formed by
galactose then condense with the phloroglucinol to form a red-colored
compound.
This forms the reduction test of carbohydrates. Fehling’s solution
Fehling’s test contains blue alkaline cupric hydroxide solution, heated with reducing
sugars gets reduced to yellow or red cuprous oxide and is precipitated.
Hence, formation of the yellow or brownish-red colored precipitate
helps in the detection of reducing sugars in the test solution.
The principle of carbohydrate fermentation states that the action of
organism on a carbohydrate substrate results in acidification of the
Fermentation test medium, detected by a pH indicator dye. A fermentation medium
consists of a basal medium containing a single carbohydrate (glucose,
lactose, sucrose, mannitol etc.)
The iodine test is based on the absorptive properties possessed by large
Iodine test polysaccharide molecules. The glucose chains in most of
polysaccharides are organized to form helices. The space between the
turns of the helix can hold small iodine molecules. This is seen with
amylase chains found in starch. Glycogen and amylopectin can also
absorb these iodine molecules on their surface. The absorptive property
of polysaccharides decreases upon heating.

NOT IN THE ACTIVITY

Moore's test This test is based on the principle that sugars boiled with concentrated
sodium hydroxide undergo polymerization to form a resinous
substance, caramel.
Reducing sugar undergo tautomerization in mildly acidic medium to
Barfoed's test form enediols. These enediols reduce cupric ions to cuprous ions that
form cuprous hydroxide. This cuprous hydroxide us converted to
cuprous oxide on heating and precipitates are formed. Its principle is
similar to Benedict’s test except the acidic environment.
Monosaccharides being strong reducing agents give this test much
early.

This test is specific for pentoses and the compounds containing ...
Tauber's Benzidine test Tauber's benzidine test: It is more specific for pentoses.
Pentoses form furfural compounds in the presence of concentrated
Bial's Orcinol Test acid. The furfural compounds formed by pentoses condense with
orcinol to form blue colored compounds.

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