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)
