Tests

There are 2 types of tests when it comes to testing for substances which are quantitative tests and qualitative
tests.

Qualitative Tests
We need to know how to carry out and interpret results for 4 biochemical tests.

Biuret test for proteins

Add biurets reagent to the test sample in a test tube.
If proteins are present then the colour of the sample will change from blue to purple.

Test for starch

Dissolve the starch in water to form the starch solution.
Add iodine solution to the starch solution.
If starch is present then there will be a colour change from brown to blue-black or purple.

Testing for reducing sugars

Dissolve the test sample in water.

Add benedict’s solution and heat the sample at 80-90 degrees for 2 minutes.
If a reducing sugar is present the colour should change from blue to brick-red.
The further the colour change, the more concentrated the reducing sugar.
Testing for non-reducing sugars
Carry out the test for reducing sugars to ensure there are none present.
Dissolve the test sample in water and add a few drops of hydrochloric acid.
Heat the solution.
Add sodium hydroxide to neutralise the solution.
Add benedict’s reagent and heat for 2 minutes.
If a non-reducing sugar is present there should be a colour change from blue to brick red.

Test strips for glucose

Test strips can also be used for glucose.
The test strip is dipped into the solution containing glucose.
A colour change in the strip will be seen depending on the concentration of glucose.

Quantitative Tests

Colorimetry

This is a colorimeter
It is used to measure the concentration
of a substance in a sample

Colorimeters measure the absorbance and transmittance of light passing through a liquid sample.
The more concentrated the colour of the sample, the higher the absorbance or the lower the transmission.

How do we find out the concentration of an unknown glucose solution?

Make several solutions of known concentrations.
Measure the absorbance and repeat for all concentrations.
Draw a table of results and plot the absorbances on a graph to obtain a calibration curve.
The calibration curve can then be used to find out the concentration of an unknown glucose solution.

Measuring the absorbance of known solutions

Carry out a benedict’s test on each solution.
Remove any precipitate by centrifuging them.
Switch on the colorimeter and add a red filter.
Use distilled water to calibrate the colorimeter
Pipette a sample of the solution to a clean cuvette and
read measure the absorbance.
Repeat for each remaining solution.
Chromatography
Chromatography allows for components to be separated in a mixture.
It consists of 2 phases: the mobile phase and the stationary phase.

The mobile phase is the solvent that moves through the paper and the stationary phase is the paper.
The distance the dye will travel is determined by the balance between the solutes attraction to the mobile phase
compared to the stationary phase.

Rf Value = Distance travelled by solute + distance travelled by solvent.