Core Practical 1: Use a semi-quantitative method with Benedict's

reagent to estimate the concentrations of reducing sugars and

with iodine solution to estimate the concentrations of starch,
using colour standards

Objectives
 To understand what is meant by a semi-quantitative test
 To be able to estimate concentrations of reducing sugars using Benedict’s reagent
 To be able to estimate concentrations of starch using iodine solution
 To develop the skills needed to plan an investigation using dilutions
Safety
 Wear eye protection.
 Avoid skin contact with Benedict’s reagent, iodine and hot solutions.
 Handle the test tubes with tongs to avoid burns.
 Do not taste the fruit juice.
Maths skills
 Solve algebraic equations.
Equipment
 eye protection  1/10 dilution of fruit juice
 test tubes, tongs and test tube rack  1 cm3 and 5 cm3 syringes
 small beakers  3 cm3 pipettes
 Benedict’s reagent  distilled water
 iodine solution  waterproof marker pen
 2% glucose solution  water bath at 60–80 °C or a large beaker
 2% starch solution of recently boiled water
 ‘unknown’ starch solution  timer or stop clock

Procedure
Procedure – Part 1: Benedict’s reagent method
1. Plan how you will use the stock 2% glucose solution to make the following five
concentrations of glucose solution: 2%, 1%, 0.5%, 1.5% and 0.25%. Write your plan in
the space provided in the Lab Book.
2. Use the waterproof pen to label six test tubes and five small beakers with the different
glucose concentrations they will contain. Label the sixth test tube ‘fruit juice’.
3. Use the syringes, the distilled water and the 2% glucose solution to create 5 cm3 of
each solution in the corresponding labelled beaker.
4. Use a clean syringe to add 2 cm3 of Benedict’s reagent to each of the six labelled test
tubes.
5. Using a clean syringe each time, add 1 cm3 of each glucose solution to the
corresponding labelled test tubes.
6. Add 1 cm3 of fruit juice to the last labelled test tube.
7. Give each tube a gentle shake to ensure the contents are mixed.

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 8. Place all six test tubes in the water bath and start the timer.
9. After 2 minutes, use tongs to remove the test tubes from the water bath and place
them in the test tube rack. Take care: the tubes will be hot.
10. Observe the tubes and record the results in a suitable table.
Procedure – Part 2: Iodine solution method
1. Plan how you will use the stock 2% starch solution to make the following five
concentrations of starch solution: 2%, 1%, 0.5%, 0.2% and 0.1%. Write your plan in the
space provided in the Lab Book.
2. Use the waterproof pen to label five test tubes and five small beakers with the different
starch concentrations they will contain. Label the sixth test tube ‘unknown’.
3. Use the syringes, the distilled water and the 2% starch solution to create 5 cm3 of each
solution in the corresponding labelled beaker.
4. Use a clean syringe to add 0.5 cm3 of iodine to each of the six labelled test tubes. Add
a further 10 cm3 of distilled water to each tube.
5. Using a clean syringe each time, add 5 cm3 of each starch solution to the
corresponding labelled test tube.
6. Add 5 cm3 of the ‘unknown’ starch solution to the sixth test tube. Compare the colour
produced in this tube with the colour of the diluted test tubes.
7. Record your results in a suitable table.
Analysis of results
1. Record your results for glucose in a suitable table.
2. If possible take a photograph of your standard solutions and keep it with your data.
3. Plot a suitable graph to display your data.
4. Record your results for starch in a suitable table.
5. If possible take a photograph of your standard solutions and keep it with your data.
6. Plot a suitable graph to display your data.
Learning tips
 When recording the results of the Benedict’s test, use the correct descriptive language.
Simply stating ‘green’ or ‘blue’ does not provide enough detail to allow comparison.
 One of the reasons for making the test semi-quantitative is to allow easier comparison,
which may be less subjective and therefore more accurate. You will be expected to
know the steps needed in the procedure and the expected results.
Questions
1. How could these investigations have been adapted to produce quantitative results?
2. Explain why the Benedict’s test is not a suitable test for measuring the total sugar
content of fruit juice.
3. Suggest one way in which the results of the Benedict’s reagent investigation could
have been made more reliable.
4. How could you evaluate the accuracy of your experimental value for the concentration
of glucose in the fruit juice?
5. Name a variable that you controlled in the iodine investigation, explain how you
controlled it and describe the possible effect on the result if it had not been controlled.

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Practical activities have been safety checked but not trialled. Users may need to adapt the risk assessment
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 Exam-style questions
1. Explain the difference between qualitative, semi-quantitative and quantitative tests.
(3)
2. Explain how you could use serial dilution to make a range of concentrations from a
stock solution of 10% fructose.
(2)
3. The graph shows a calibration curve obtained by measuring the absorbance of light of
wavelength 650 nm in a colorimeter, through a solution of starch reacted with dilute
iodine/KI solution.

(a) Suggest why light of wavelength 650 nm was used in the colorimeter.
(2)
(b) Explain why a blank – a cuvette containing distilled water and dilute iodine/KI
solution – is used to set the absorbance of the colorimeter to zero.
(1)
(c) Explain why a scratched cuvette should not be used.
(1)
(d) Dilute iodine/KI solution was added to a starch solution of unknown concentration
and mixed. The resulting solution was placed in a cuvette in a colorimeter and the
absorbance was read as 0.75. What was the concentration of starch in this
solution?
(1)
(e) Express the concentration of starch read from the graph in (d) as:
(i) a percentage
(ii) g per L
(2)
4. Explain what is meant by a ‘reducing sugar’.
(3)
5. Starch is a polymer of glucose. Name the type of reaction involved when two glucose
molecules join together by a 1,4 glycosidic bond.
(1)

© Pearson Education Ltd 2018. Copying permitted for purchasing institution only. This material is not copyright free.
Practical activities have been safety checked but not trialled. Users may need to adapt the risk assessment
information to local circumstances. This document may have been altered from the original. 3
© Pearson Education Ltd 2018. Copying permitted for purchasing institution only. This material is not copyright free.
Practical activities have been safety checked but not trialled. Users may need to adapt the risk assessment
information to local circumstances. This document may have been altered from the original. 4