Biochemical Tests Flashcards
Reducing sugars
Benedict’s
Reducing sugars test method
Add Benedict’s reagent (which is blue as it contains copper (II) sulfate ions) to a sample solution in a test tube
Heat the test tube in a water bath or beaker of water that has been brought to a boil for a few minutes
If a reducing sugar is present, a coloured precipitate will form as copper (II) sulfate is reduced to copper (I) oxide which is insoluble in water
It is important that an excess of Benedict’s solution is used so that there is more than enough copper (II) sulfate present to react with any sugar present
A positive test result is a colour change somewhere along a colour scale from blue (no reducing sugar), through green, yellow and orange (low to medium concentration of reducing sugar) to brown/brick-red (a high concentration of reducing sugar)
This test is semi-quantitative as the degree of the colour change can give an indication of how much (the concentration of) reducing sugar present
Non-reducing sugars
Hydrochloric acid & Benedicts
Non-reducing sugars test method
Add dilute hydrochloric acid to the sample and heat in a water bath that has been brought to the boil
Neutralise the solution with sodium hydrogencarbonate
Use a suitable indicator (such as red litmus paper) to identify when the solution has been neutralised, and then add a little more sodium hydrogencarbonate as the conditions need to be slightly alkaline for the Benedict’s test to work
Then carry out the Benedict’s test as normal; add Benedict’s reagent to the sample and heat in a water bath that has been boiled – if a colour change occurs, a reducing sugar is present
Starch
Iodine
Starch test method
To test for the presence of starch in a sample, add a few drops of orange/brown iodine in potassium iodide solution to the sample
The iodine is in potassium iodide solution as iodine is insoluble in water
If starch is present, iodide ions in the solution interact with the centre of starch molecules, producing a complex with a distinctive blue-black colour
This test is useful in experiments for showing that starch in a sample has been digested by enzymes
Protein
Biuret’s
Protein test method
Add Biuret’s reagent to sample
If protein present: colour change from blue to purple
Lipids
Emulsion test
Lipids test method
Dissolve sample in ethanol
Pour solution slowly into water
If lipids present: white emulsion will form