Core Practical 1– Wednesday, December 13, 2023

Benedict’s reagents and reducing sugars

Learning Objectives:

https://www.youtube.com/watch?v=C765WC8htwY

• AS Specification reference no.1.3

Do now: 10 mins
‘Describe and draw’ in pairs.
• Each student is given a different card. One student describes the molecule on the
card and the second must draw it. They then swap over cards.
• Allow a maximum of 2 mins for each drawing
• The first pair to get all their drawing completed in 10 mins wins!
- You will need cards displaying the formulae of different monosaccharides and disaccharides.
 CP1 - 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 EQUIPMENT
● eye protection
● To understand what is meant by a semi- ● test tubes, tongs and test tube rack
quantitative test ● small beakers
● To be able to estimate concentrations of ● Benedict’s reagent
● iodine solution
reducing sugars using Benedict’s reagent ● 2% glucose solution
● To be able to estimate concentrations of ● 2% starch solution
starch using iodine solution ● ‘unknown’ starch solution
● 1/10 dilution of fruit juice 1 cm3 and 5 cm3 syringes
● To develop the skills needed to plan an ● 3 cm3 pipettes
investigation using dilutions ● distilled water
● waterproof marker pen
● water bath at 60–80 °C or a large beaker of recently boiled
water
● timer or stop clock
 Core practical 1

WARM UP QUESTIONS (iGCSE RECALL)

REMOVE BOX TO REVEAL ANSWER!

 What are we testing for when we use Benedict’s
solution?
• Reducing sugars

• ALL monosaccharides and

some disaccharides are
Reducing Sugars

Can you name reducing sugars?

 What is a reducing sugar?
https://www.youtube.com/watch?v=d6tHWPW5WLM

Make notes from the video – stop at 1.03

• A substance that provides electrons to carry out
reduction
• Benedict’s reagent contains copper II sulphate.
• The copper ions are reduced by electrons from
the CO group found in specific sugars.
• This produces copper I oxide (insoluble, red
precipitate)
 What if you have a non reducing sugar?

• It must first be hydrolysed into its monosaccharide components:

• First test with Benedict’s reagent

• If no colour change:
• Add 2cm³ of food sample to 2cm³ of dilute HCl
• Place in gently boiling water bath for 5 mins
• Slowly add some sodium hydrogen carbonate solution until neutralized
and allow the solution to cool down
• Re-test using Benedict’s reagent

• Alternatively, use the enzyme sucrase to hydrolyze

Test : Benedicts test for reducing and non-reducing sugars

 Carry out a test for glucose and sucrose before and after adding HCl and
heating and then neutralising with sodium hydrogen carbonate.
 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 a detailed explanation of your observations.

Use as many keywords as possible in your explanation of the

breakdown/hydrolysis of sucrose to fructose and glucose

https://www.youtube.com/watch?v=QU0VBcHnQOk

Equipment / materials
Sucrose, hydrochloric acid, sodium hydrogen carbonate, water, Bunsen burner, beaker, water.
 Making a
serial dilution
 Semi-quantitative
assessment of reducing
sugar content

No reducing sugar High

https://www.youtube.com/watch?v=ssgq1QbgJrM

Produce a range of colour standards with known concentrations of glucose and

these can be used to compare the result of a test carried out with an equal volume of
an unknown solution e.g. orange juice
HOW DO I MAKE THE EXPERIMENT QUANTITAIVE?
By using a colorimeter to
measure the amount of light being
absorbed by the solution.
Remember to blank the
colorimeter with the solution of
Benedicts reagent.
Why is this necessary?

A graph can then be plotted and used to determine the concentration of glucose
in an unknown solution. This is called a calibration curve.
 • Use solutions of known concentrations to produce a set of values.
Calibration • Plot a calibration curve

curve
• Find a value for the unknown concentration
• Draw intercepts on the graph to determine the concentration of the unknown solution
• Using the calibration curve, estimate the concentration of reducing sugar in the orange juice

Concentration of Colorimeter reading

reducing sugar (Absorbance)
solution (%)

4.00 0.05
2.00 0.25
1.00 0.65
0.50 0.96
0.25 1.15
Orange juice?? 0.12

• For low concentrations of glucose a positive Benedicts reagent result ranges from green through to brick red. By
comparing the colour of the sample solution with the colour of a standard solution, the glucose concentration of
the sample can be estimated.
EXAM QUESTION 1 5 MINUTES

Benedict’s test was carried out on five food samples. The results are shown below

Sample Colour of solution

A Yellowish brown
B Green
C Red
D Dark brown
E Yellowish green

A. List the letters in sequence of the increasing amount of reducing sugar in each sample.
B. Suggest a way, other than comparing colour changes, in which different concentrations of
reducing sugar could be estimated.
C. Explain why it is not possible to distinguish between very concentrated samples, even when
their concentrations are different
EXAM QUESTION 1 - answers

A. List the letters in sequence of the increasing amount of reducing sugar in each sample.
BEADC
B. Suggest a way, other than comparing colour changes, in which different concentrations of
reducing sugar could be estimated.
Dry the precipitate in each sample and weigh it. The heavier the precipitate the more reducing
sugar is present.

C. Explain why it is not possible to distinguish between very concentrated samples, even
when their concentrations are different
As soon as all the copper II sulphate has been reduced to copper I oxide, further amounts of
reducing sugar would not make a difference.
Test 2 : Benedicts test for reducing and non-reducing sugars

Students carry out a test for glucose and sucrose before and after adding HCl
and heating and then neutralising with sodium hydrogen carbonate.

Students write a detailed explanation of their observations.

Students should use as many keywords as possible in their

explanation of the breakdown/hydrolysis of sucrose to fructose and
glucose
https://www.youtube.com/watch?v=QU0VBcHnQOk

Equipment / materials
Sucrose, hydrochloric acid, sodium hydrogen carbonate, water, Bunsen burner, beaker, water.
 https://www.youtube.com/watch?v=d6tHWPW5WLM

(26) A Level Biology Revision "Using a colorimeter to quantify gl

ucose 2"
– YouTube
 CP1 - 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.

EQUIPMENT
OBJECTIVES ● eye protection
● test tubes, tongs and test tube rack
● To understand what is meant by a semi-
● small beakers
quantitative test ● Benedict’s reagent
● To be able to estimate concentrations of ● iodine solution
● 2% glucose solution
reducing sugars using Benedict’s reagent
● 2% starch solution
● To be able to estimate concentrations of ● ‘unknown’ starch solution
starch using iodine solution ● 1/10 dilution of fruit juice 1 cm3 and 5 cm3 syringes
● 3 cm3 pipettes
● To develop the skills needed to plan an
● distilled water
investigation using dilutions ● waterproof marker pen
● water bath at 60–80 °C or a large beaker of recently boiled
water
● timer or stop clock
 Serial dilutions
Part 2: Iodine solution method
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 your book.
 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.
 Error bars (Excel)
Error bars are expressed as the percentage of possible error amounts for each data point in a two-dimensional chart. They help
indicate whether there are significant differences between two variables that are being compared. Standard error bars represent
the standard deviation of the mean. Calculating standard errors and drawing the bars manually can be tedious. Microsoft Excel
makes this task easier with it Standard Error bar feature. Learn how to add and customize standard error bars in charts using
Microsoft Excel.

Step 1 - Open a new workbook in Microsoft Excel. Enter data points in cells down two columns. For example, if we want to plot
the number of students that earned a certain average exam grade, we would list the number of students down column A and the
average grade in column B.

Step 2 - Highlight the data in both columns. Go to the "Insert" tab and choose a chart type. For example, we can choose a scatter
chart to plot the data.

Step 3 - Click on a data point to select all of the data points in the chart, or click twice on a data point to only select that data
point. You can have standard error bars for one data point or all of the data points.

Step 4 - Go to the "Layout" tab under Chart Tools and click on "Error Bars." Select "Error Bars with Standard Error."

Step 5 - Click on the "Error Bars" menu and select "More Error Bar Options." You can change the direction of the error bars, the
end style, and the error amount. The line color, style and shadow can also be customized.
Helpful links

https://www.youtube.com/watch?v=C765WC8htwY