BIOCHEMISTRY (LAB) - MED223
CARBOHYDRATES, PROTEINS, LIPIDS
CARBOHYDRATES
● Contains Carbon, Hydrogen, Oxygen
● Oxidation of sugars provides energy for
organisms to carry out their life
processes.
● The highest yield of energy from
carbohydrates occurs when sugars are
completely oxidized to CO2 and H2O in
the aerobic process.
● Reduction of CO2 and H2O to form
sugars, a process that takes place in
photosynthesis.
ALDEHYDE GROUPS
● Can be oxidized to give the carboxyl
group that is the characteristic of acids,
and this reaction is the basis of a test for
the presence of aldoses.
● Aldoses are called reducing sugars
because of this type of reaction
● Ketoses can also be reducing sugars
because they isomerize to aldoses.
● Lactone
○ The compound produced by
oxidation of an aldose
○ A cyclic ester linking the carboxyl
group and one of the sugar
alcohols.
Monosaccharides
● Simple sugars; building blocks of
carbohydrates
● Glucose is an example of a carbohydrate
monomer or monosaccharide.
● Other examples of monosaccharides
include mannose, galactose, fructose,
etc.
● are those carbohydrates that cannot be
hydrolyzed further to give simpler units
of polyhydroxy aldehyde or ketone.
Disaccharides
● Two monosaccharides combined
● Sucrose, Lactose, Maltose, etc.
● Maltose and Lactose (also known as milk
sugar) are the other two important
disaccharides.
Oligosaccharides
● Formed when a few monosaccharides are
linked together
● Carbohydrates formed by the
condensation of 2-9 monomers
● trioses, pentoses, hexoses are all
oligosaccharides.
Polysaccharides
● Formed when many monosaccharides
are bonded together
● Complex carbohydrates
● are referred to as starchy foods
● It is a polymer of α glucose and consists
of two components-Amylose and
Amylopectin.
● starch, glycogen, cellulose, etc.
Reducing sugars
● Those sugars which act as reducing
agents
● They reduce Fehling solution and Tollen's
reagent.
● All monosaccharides and disaccharides,
excluding sucrose, are reducing sugars.
● It can be further reduced to alcohol.
Non-reducing sugars
● is not reduced by a weak oxidizing agent
(one that oxidizes aldehydes but not
alcohols, such as Tollen's reagent).
● do not have an —OH group attached to
the anomeric carbon so they cannot
reduce other compounds.
BIOCHEMISTRY (LAB) - MED223
CARBOHYDRATES, PROTEINS, LIPIDS

Molisch’s Test
Sucrose
● is used to check for the presence of
carbohydrates in a given analyte. Starch
● a purple or a purplish-red ring at the
point of contact between the H2SO4 and Manose
the analyte + Molisch’s reagent mixture
confirms the presence of carbohydrates MOLISCH’S TEST
in the analyte. Presence of
SOLUTIONS Observation
Carbohydrates
Fehling’s Test
Red to Purple
● used for the estimation or detection of Glucose ✓
Ring
reducing sugars and non-reducing
Red to Purple
sugars Lactose ✓
Ring
Benedict’s Test Blue to Purple
Sucrose ✓
● is used to check the presence of Ring
reducing sugars. Starch Purple Ring ✓
● formation of brick-red precipitate that
Reducing/
indicates a positive test. Solutions Benedict’s Fehling’s
Non-reducing
Tollen’s Test Glucose + + Reducing
● useful method to distinguish between Lactose + + Reducing
aldehydes and ketones. Sucrose - - Non-reducing
● Tollens test is commonly referred to as Starch - - Non-reducing
the Silver Mirror test due to the formation
TOLLEN’S TEST
of this layer of metallic silver on the test
tube. Aldehyde/
SOLUTIONS Observation
Ketone
● Silver Mirror positive result
Glucose Silver Mirror Aldehyde
Iodine Test
Lactose Silver Mirror Aldehyde
● can help to distinguish starch from
Sucrose No Silver Mirror Ketone
monosaccharides, disaccharides, and Starch No Silver Mirror Ketone
other polysaccharides.
IODINE’S TEST
MOLECULAR STRUCTURE
SUGARS
(ring form) SOLUTIONS Observation
Produced an Amber/Orange color
Glucose
which means no reaction.
Produced an Amber/Orange color
Glucose Lactose
which means no reaction.
Produced an Amber/Orange color
Sucrose
which means no reaction.
Produced a Blue color which
Lactose Starch
means a reaction occurred.
BIOCHEMISTRY (LAB) - MED223
CARBOHYDRATES, PROTEINS, LIPIDS

PROTEINS MILLON’S TEST

● Proteins are polymers of amino acids, ● Phenolic group of tyrosine of proteins
with each amino acid residue joined by react with mercuric sulfate in the
peptide bonds presence of sodium nitrite and sulfuric
● Proteins can be broken down acid to give red color
(hydrolyzed) to their constituent amino ● brick red color solution confirms the
acids by a variety of methods, and the presence of proteins
earliest studies of proteins naturally NINHYDRIN TEST
focused on the free amino acids derived ● The pyridine solution of ninhydrin gives
from them. coloured solution from deep blue to
● Residue reflects the loss of the elements violet pink, sometimes even red color in
of water when one amino acid is joined to some cases.
another. ● violet color solution confirms the
● AMINO ACIDS presence of proteins
- building blocks of proteins
LIPIDS
- are organic compounds containing the
basic amino groups (-NH2) and carboxyl SOLUBILITY TEST
groups (-COOH). SAMPLE WATER ALCOHOL CHLOROFORM
● POLYPEPTIDES - long chain of monomers Miscible upon
Desi Ghee Immiscible Miscible
that are held by peptide bonds to form heating
protein Vegetable Miscible upon
Immiscible Miscible
Oil heating
● PEPTIDE BONDS - are amide between
Refined Miscible upon
that carboxyl of one amino acid and the Immiscible Miscible
Oil heating
a-amino acid of another.
● DENATURED PROTEINS - are unfolded TRANSLUCENT SPOT TEST
and disorganized proteins in its
SAMPLE OBSERVATION
secondary and tertiary structure without
hydrolysis of peptide bonds. Translucent spot appears on filter
Desi Ghee
paper.
BIURET TEST
Vegetable Translucent spot appears on filter
● Identification of proteins Oil paper.
● Biuret reagent in the presence of copper
ions forms a violet color complex. Translucent spot appears on filter
Refined Oil
paper.
● Violet color - positive reaction
XANTHOPROTEIC TEST
● yellow or orange color
● Concentrated nitric acid is used for
nitration
BIOCHEMISTRY (LAB) - MED223
CARBOHYDRATES, PROTEINS, LIPIDS

ACROLEIN TEST

SAMPLE OBSERVATION

A pungent irritating color of

Desi Ghee
Acrolein.

Vegetable A pungent irritating color of

Oil Acrolein.

A pungent irritating color of

Refined Oil
Acrolein.

BAUDOUIN TEST

SAMPLE OBSERVATION

Desi Ghee No rose-red color appears.

Vegetable
Rose-red color appears.
Oil

HUBLE’S TEST

SAMPLE OBSERVATION

Desi Ghee Violet color does not fade away.

Vegetable
Violet color fades away.
Oil