AQA A Level Biology Biochemical Tests
What are reducing sugars?
ALL MONOSACCHARIDES + some disaccharides like MALTOSE AND LACTOSE.
Reducing sugars can donate electrons (the carbonyl group becomes oxidised), so the sugars become the reducing agent.
So reducing sugars can be detected using Benedict’s test as they reduce the soluble copper sulphate into insoluble brick-red copper oxide.
Describe the Benedict’s test for reducing sugar?
Add the Benedict’s reagent (which is blue as it contains copper (II) sulfate ions) to the sample in a test tube.
Heat the sample with excess benedict’s reagent in a gently boiling water bath for 5 minutes.
If it stays blue, there is no reducing sugar present. If it’s brick red, an insoluble precipitate forms then a reducing sugar is present.
Note- Benedict’s reagent is copper (II) sulphate; in the presence of a reducing sugar, copper (II) sulphate is reduced to copper (I) oxide.
In increasing amount of reducing sugar the colour goes
blue>green>yellow>orange>red.
What are non-reducing sugars?
Some types of disaccharides for example sucrose but NO monosaccharides.
Non-reducing sugars cannot donate electrons, therefore they cannot be oxidised.
To be detected non-reducing sugars must first be hydrolysed to break the disaccharide into its two monosaccharides before a Benedict’s test can be carried out.
Describe the Benedict’s test for non-reducing sugars?
Add a few drops of dilute hydrochloric acid (hydrolyse sugar into its constituent reducing sugar) into the sample.
Heat the sample in a boiling water bath
Neutralise the solution with sodium bicarbonate.
Add Benedict’s reagent and heat again
If a non-reducing sugar is present a green/yellow/orange or red predicate will form.
The reason is
• The addition of acid will hydrolyse any glycosidic bonds present in any carbohydrate molecules.
• The resulting monosaccharides left will have an aldehyde or ketone functional group that can donate electrons to copper (II) sulfate (reducing the copper), allowing a precipitate to form.
Describe a test for starch?
Add iodine dissolved in potassium iodide solution to the solution and shake/stir
If starch is present → colour change from orange/brown to blue/black.
How do you use chromatography to detect monosaccharides?
Mixtures containing coloured molecules, such as ink or chlorophyll, do not have to be stained as they are already coloured.
Mixtures of colourless molecules, such as a mixture of monosaccharides, have to be stained first.
A spot of the stained monosaccharide sample mixture is placed on a line at the bottom of the chromatography paper.
Spots of known standard solutions of different monosaccharides are then placed on the line beside the sample spot.
The chromatography paper is then suspended in a solvent.
As the solvent travels up through the chromatography paper, the different monosaccharides within the mixture separate out at different distances from the line.
The unknown monosaccharides can then be identified by comparing and matching them with the chromatograms of the known standard solutions of different monosaccharides.
If a spot from the monosaccharide sample mixture is at the same distance from the line as a spot from one of the known standard solutions, then the mixture must contain this monosaccharide.
What are reducing sugars?
ALL MONOSACCHARIDES + some disaccharides like MALTOSE AND LACTOSE.
Reducing sugars can donate electrons (the carbonyl group becomes oxidised), so the sugars become the reducing agent.
So reducing sugars can be detected using Benedict’s test as they reduce the soluble copper sulphate into insoluble brick-red copper oxide.
Describe the Benedict’s test for reducing sugar?
Add the Benedict’s reagent (which is blue as it contains copper (II) sulfate ions) to the sample in a test tube.
Heat the sample with excess benedict’s reagent in a gently boiling water bath for 5 minutes.
If it stays blue, there is no reducing sugar present. If it’s brick red, an insoluble precipitate forms then a reducing sugar is present.
Note- Benedict’s reagent is copper (II) sulphate; in the presence of a reducing sugar, copper (II) sulphate is reduced to copper (I) oxide.
In increasing amount of reducing sugar the colour goes
blue>green>yellow>orange>red.
What are non-reducing sugars?
Some types of disaccharides for example sucrose but NO monosaccharides.
Non-reducing sugars cannot donate electrons, therefore they cannot be oxidised.
To be detected non-reducing sugars must first be hydrolysed to break the disaccharide into its two monosaccharides before a Benedict’s test can be carried out.
Describe the Benedict’s test for non-reducing sugars?
Add a few drops of dilute hydrochloric acid (hydrolyse sugar into its constituent reducing sugar) into the sample.
Heat the sample in a boiling water bath
Neutralise the solution with sodium bicarbonate.
Add Benedict’s reagent and heat again
If a non-reducing sugar is present a green/yellow/orange or red predicate will form.
The reason is
• The addition of acid will hydrolyse any glycosidic bonds present in any carbohydrate molecules.
• The resulting monosaccharides left will have an aldehyde or ketone functional group that can donate electrons to copper (II) sulfate (reducing the copper), allowing a precipitate to form.
Describe a test for starch?
Add iodine dissolved in potassium iodide solution to the solution and shake/stir
If starch is present → colour change from orange/brown to blue/black.
How do you use chromatography to detect monosaccharides?
Mixtures containing coloured molecules, such as ink or chlorophyll, do not have to be stained as they are already coloured.
Mixtures of colourless molecules, such as a mixture of monosaccharides, have to be stained first.
A spot of the stained monosaccharide sample mixture is placed on a line at the bottom of the chromatography paper.
Spots of known standard solutions of different monosaccharides are then placed on the line beside the sample spot.
The chromatography paper is then suspended in a solvent.
As the solvent travels up through the chromatography paper, the different monosaccharides within the mixture separate out at different distances from the line.
The unknown monosaccharides can then be identified by comparing and matching them with the chromatograms of the known standard solutions of different monosaccharides.
If a spot from the monosaccharide sample mixture is at the same distance from the line as a spot from one of the known standard solutions, then the mixture must contain this monosaccharide.