DEPARTMENT OF BIOLOGY/BIOTECHNOLOGY,

DAVID UMAHI FEDERAL UIVERSITY OF HEALTH SCIENCES,

UBURU

FOOD TEST
Presented By

NWEKE AUGUSTINE
OGBU C. A
UGADU JOHN
 Food Tests: Identifying
TOPIC 1 Macronutrients
01 INTRODUCTION
Food tests are essential laboratory procedures used to
identify the presence of various macronutrients
(carbohydrates, proteins, lipids and nucleic acids) in
food samples.
These tests rely on specific chemical reactions that
produce observable changes, such as color changes or
precipitate formation, indicating the presence or
absence of a particular nutrient.
02 BELOW ARE THE MACROMOLECULES AND
THEIR BUILDING BLOCKS
Macromolecule Building Block
Carbohydrates Monosaccharide
Protein Amino Acids

Lipids Glycerol + Fatty Acids

Nucleic Nucleotides (DNA and RNA)
3 Tests For Carbohydrate
 BENEDICT'S TEST FOR REDUCING
SUGARS
(E.G., GLUCOSE, FRUCTOSE)

4
05 Introduction

Benedict's test is a chemical test used to detect the

presence of reducing sugars such as glucose, fructose,
and lactose.
It is based on the reduction of cupric ions (Cu^2+) to
cuprous ions (Cu^+) in an alkaline solution, forming a
colored precipitate of copper(I) oxide.
06 Principle

Benedict's reagent consists of copper(II) sulfate, sodium

citrate, and sodium carbonate in an alkaline solution.
When heated with a reducing sugar, the blue cupric ions
(Cu^2+) are reduced to red cuprous oxide (Cu_2O) or
yellow cuprous hydroxide (Cu_2(OH)_2), depending on
the concentration of reducing sugar present.
07 Materials:
Beaker,
Pipette,
Test Tube,
Water,
Water Bath
 Reagents:
08
Copper sulphate
Sodium citrate
Sodium carbonate
09 Procedures:
Pipette 1ml of carbohydrate solution into a test tube
Add 5ml of benedict’s reagent
Shake to mix thoroughly
Place the test tube in a boiling water bath for 3mins
Observe the colour changes
10 Indication:
Note the colour of the precipitate at the base of the test
tube.
11 Results:

Negative: Blue solution (no reducing sugar present).

Positive: Green, yellow, orange, or red precipitate
(indicating the presence of reducing sugars, with the
color intensity reflecting the concentration).
Observations:

Blue colour = no sugar or 0% or negative

Green colour = 0.5% sugar or +
Yellow colour = 1% sugar or ++
Orange colour = 1.5% sugar or +++
Brick red colour = 2% sugar or ++++
Precautions:

Wash the apparatus before and after experiment

Measure the correct volume of the reagent
Avoid prolonged heating of the test tube
Carefully handle all the chemicals.
FEHLINGS TEST FOR
REDUCING SUGAR
Introduction:

Fehling's test is a chemical test used to detect the

presence of reducing sugars such as glucose, fructose,
and lactose.
It is based on the reduction of cupric ions (Cu^2+) to
cuprous ions (Cu^+), forming a brick-red precipitate
of copper(I) oxide.
Principle:

The principle behind Fehling's test (same as benedicts

test) lies in the ability of reducing sugars to reduce
cupric ions in an alkaline solution.
In Fehling's solution, there are two separate solutions:
Fehling's A (aqueous copper(II) sulfate) and Fehling's
B (potassium sodium tartrate and sodium hydroxide).
When the two solutions are mixed in equal volumes
and heated with a reducing sugar, the blue cupric ions
(Cu^2+) are reduced to insoluble red cuprous oxide
(Cu_2O).
Materials:

Carbohydrate solution,
beaker,
test tubes,
pipette,
water,
water bath.
Reagents:

Fehling’s solution A
Fehling’s solution B
Procedures:

Add 2ml of carbohydrate/sugar solution into a test

tube
Add 1ml each of Fehling A and B solution into the
same test tube
Boil the mixture for 5mins
Indication:

The production of yellow or brownish-red precipitate

of Cuprous oxide indicates the presence of sugars in
the sample.
Result:
Reducing sugar is present.
Precautions:

Wash the apparatus before and after experiment

Pipette the correct volume of the solutions
Mix and boil the sample carefully.
SELIWANOFF’S TEST FOR
KETO SUGARS
Introduction

Seliwanoff’s test is used to differentiate between keto

sugars and aldose sugar.
Aldose: Is a type of monosaccharide or simple
carbohydrate that contains an aldehyde in its structure.
Ketose: are those monosaccharide's that contain
ketone functional group in their structure.
Principle:

Carbohydrates are converted to furfural derivatives by

dilute HCL.
Only furfural derivatives of keto-hexoses condense
with resorcinol to form a cherry red complex.
Materials

Carbohydrate solution
Beaker
Test tubes
Pipette
Water
Water bath
Reagents

Seliwanoff’s reagent
Resorcinol
Concentrated HCL
Procedures

Pour 1 ml of carbohydrate solution (original solution)

into a test tube
Add 5 ml of seliwanoff’s reagent
Heat the test tube in a boiling water bath for 30
seconds
Observe the colour change.
 Result

Positive result (keto present): A rapid formation of a cherry-

red color indicates the presence of an ketose sugar (like
fructose)

Negative result (Aldose Present): A slow formation of faint

pink or light yellow colour indicates the presence of an aldose
sugar.
IODINE TEST FOR
POLYSACCHARIDES
STARCH IODINE TEST

Introduction:
The iodine test is a chemical test used to detect the
presence of polysaccharides, particularly starch, in a
given sample.
It relies on the formation of a colored complex
between starch and iodine, which produces a
characteristic blue-black coloration.
Principle

The principle behind the iodine test lies in the ability

of starch to form a complex with iodine molecules.
Iodine molecules penetrate the helical structure of
starch molecules and interact with the hydroxyl groups
present in the glucose units.
This interaction results in the formation of an
inclusion complex, leading to the characteristic blue-
black coloration.
Procedure

1. Prepare a dilute solution of iodine (typically iodine

dissolved in potassium iodide solution).
2. Add a small amount of the sample (containing the
polysaccharide) to a test tube or onto a white surface.
3. Add a few drops of the iodine solution to the
sample.
4. Observe for the formation of a blue-black
coloration.
Interpretation

- Positive Result: The formation of a blue-black

coloration indicates the presence of polysaccharides,
particularly starch, in the sample.
- Negative Result: No color change or only a slight
color change indicates the absence of polysaccharides.
Precautions

- Use a dilute solution of iodine to avoid excessive

staining.
- Ensure that the sample is properly dispersed in the
solvent to facilitate interaction with iodine molecules.
- Dispose of waste properly, as iodine solutions can be
hazardous and stain surfaces.
Conclusion

The iodine test is a simple and effective method for

the detection of starch and other polysaccharides in
various samples.
By understanding the principles, procedure, and
interpretation of the test, one can utilize it for
qualitative analysis in research, quality control, and
educational settings.
Laboratory Tests for
Protein
Introduction to Protein Testing

Proteins are essential macromolecules involved in

numerous biological processes.
Laboratory tests for protein are crucial in clinical
diagnostics, food analysis, and research.
These tests can determine the presence, concentration,
and type of proteins in various samples.
 Principles of Protein Detection

Protein detection methods rely on the chemical properties

of amino acids and peptide bonds.
These methods can be classified based on the principle of
detection:
Colorimetric assays: Detect changes in color due to
protein-reagent interactions.
Principles of Protein Detection cont’d

Spectrophotometric Assays: Measure the absorbance

or transmittance of light by proteins.
Immunological Assays: Use antibodies to detect
specific proteins.
Electrophoresis: separates proteins based on size and
charge.
Common Laboratory Tests for Protein

Millon's Test
Biuret Test
MILLON’S TEST
(for detection of amino acid “tryosine”)
 Principles
The mercurous and mercuric nitrate reacts with
the hydroxybenzene radicals (phenols) forming a
red coloured compound.
In other words, Millon’s reagent reacts with
phenolic group of tyrosine to form mercuric
fumarate which gives pinkish or red coloured
compound.
 Materials:
Protein solution (egg albumin),
beaker,
pipette,
test tube,
water,
water bath
Reagent

Millon’s reagent
 Procedures:

Pipette 2ml of protein solution into a test tube

Add few drops of the Millon’s reagent
Boil the mixture of the solution for about
30sec.
Observe the colour changes
 Indication:
You will observe that the solution turns red or that
a red precipitate is produced.

The protein on the addition of Millon’s reagent,

form a white precipitate first due to denaturation
of proteins by mercury salts, which upon heating
turns red.
 Precautions:
Wash the apparatus before and after experiment
Test tubes should be free of impurities before pouring
chemicals
Carefully measure the chemicals before adding it to the test
tubes
Carefully handle the test tubes near the fire burner
Avoid prolonged heating of the test tube as this might give
false result.
 Biuret Test
Principles
In the presence of alkaline medium, cupric ions
given CUSO4 react with peptide linkage to form
the bluish violet coloured complex. The peptide
nitrogen atoms form a coordination complex with
the cupric ions. The intensity of the colour depends
on the amount of the proteins present.
Materials

Protein solution, test tubes, and pipettes

Reagents
Copper sulphate CUSO4

Sodium hydroxide (NaOH)

 Procedure
Pipette 2ml of protein solution into a test tube

Add two drops of 2% cupper sulphate solution

Add 1ml of 5% sodium hydroxide solution

Mix thoroughly and observe the colour changes

 Result
Positive Result: A colour change from the initial blue of the
Biuret reagent to a violet or purple colour indicates the presence
of proteins.
Negative Result: if the solution remains blue, it indicates the
absence of proteins or a very low concentration of them.

Blue= No protein
Purple= Protein present
 LABORATORY TEST FOR LIPIDS

Sudan III for lipids

Ethanol Emulsion test for fats and oil

Grease spot test

 SUDAN III TEST FOR LIPIDS

The Sudan III test is a simple test for detecting the

presence of lipids (fats and oil) in a sample

It relies on the principle that Sudan III, a fat-

soluble dye will dissolve in lipids, staining them
red or orange.
 Principle
Sudan III is a lysochrome dye, meaning it is soluble in
lipids but not in water.

When Sudan III is added to a sample containing lipids,

the dye dissolves in the lipid molecules.
 Materials

Test tubes,
oil,
water,
Sudan III
 Procedures
Add 2mls of oil to 2mls of water in a test tube

Add a few drops of Sudan III and shake

 Observation
Oils are stained red with Sudan III.

Since they are less dense than water, they separate out as
a red layer or globule on the water surface in the test tube
 Result
Positive: If lipids are present, the Sudan III dye which is
a red fat soluble dye will dissolve in the lipid. This will
result in the lipid being stained a red or orange colour.
This indicates the sample contains lipids.

Negative result: if lipids are absent, the Sudan III dye

will not dissolve and there will be no significant red or
orange coloration. The dye will remain separate from the
sample.
 ETHANOL EMULSION TEST FOR FATS
AND OIL
Principles
Lipids are on-polar organic compounds. Hence, they
are soluble in organic solvents such ass ethanol, but
insoluble in water.
 Materials
Test tubes, lipid sample, water and ethanol
 Procedures
Add a few drops of the lipid sample to a dry test tube

Add 2 ml of ethanol and shake thoroughly

Add 2 ml of deionized water

 Result
Positive: Cloudy white emulsion

The key indicator is the formation of a milky-white or cloudy

emulsion.

Negative: the mixture will remain clear or may exhibit only

slight turbidity.
 GREASE SPOT TEST

Principles
If a drop of liquid (such as alcohol, water or oil) falls on a paper
it makes a mark which is bright and translucent when the paper is
held up to the light. With some liquids, the mark disappears as
soon as the liquid evaporates. Alcohol takes a few second to
disappear, water takes about a minute, but oil remains forming a
permanent grease spot.
 Materials
Test tubes, test tube rack, pipette, filter papers,
alcohol, water, oil, food sample
 Procedures

Draw up some of the liquid using a pipette

Make a drop of the liquid onto a filter paper
Using a clean pipette, draw similar quantity of alcohol and
make a drop on clean part of the filter paper or a new paper
Repeat step 3 with water (alcohol and water serves as control
samples)