PRACTICAL 11

Required practical Production of a dilution series of a glucose solution and use of

colorimetric techniques to produce a calibration curve with which to
identify the concentration of glucose in an unknown ‘urine’ sample
Apparatus and b. Use appropriate instrumentation to record quantitative measurements,
techniques covered such as a colorimeter
c. Use laboratory glassware apparatus for a variety of experimental
(Not full statements) techniques to include serial dilutions
f. Use qualitative reagents to identify biological molecules

Indicative apparatus glucose solution, distilled water, Clinistix, Benedicts solution, graduated
pipettes and fillers, test tubes, water bath, colorimeter and cuvettes

Amount of choice
Increasing independence
Least choice Some choice Many choices Full investigation
Teacher chooses Teacher Student decides Student is presented with
the concentration provides on range of the urine of suspected
of standard concentration of glucose diabetic.
glucose solution standard concentrations for Student researches
and values for glucose solution calibration curve. methods for finding out the
serial dilution. but students Students produce concentration of glucose in
Volumes of decide on values dilutions and after the urine, then chooses
glucose solution for serial reacting with equipment and materials,
and water are dilution. Benedicts reagent justifying all choices.
given for serial Students produce a
dilution. Students produce calibration curve.
produce dilutions dilutions and Unknown ‘urine’
and after reacting after reacting sample is reacted
with Benedicts with Benedicts with Benedicts
reagent produce a reagent produce reagent and the
calibration curve. a calibration value of glucose
Unknown ‘urine’ curve. Unknown concentration is
sample is reacted ‘urine’ sample is read from the
with Benedicts reacted with calibration curve.
reagent and the Benedicts Teacher provides
value of glucose reagent and the outline method
concentration is value of glucose only.
read from the concentration is
calibration curve. read from the
Experiments fully calibration
specified in terms curve.
of equipment and Experiment
method. probably fully
specified by
teacher.
Opportunities for observation and assessment of competencies
 Students  Students  Students  Students follow a
Follow written
follow written follow written follow an outline method they have
procedures
method. method. method researched.
 Applies
investigative  Students must
Students
approaches  Students  Students must choose an appropriate
must correctly
and methods must correctly use correctly use the approach, equipment and
use the
when using the appropriate appropriate techniques and, identify
appropriate
instruments equipment. equipment. correct variables for
equipment.
and measurement and control.
equipment
Safely uses a
 Students
range of  Students must  Students must  Students must carry
minimise risks
practical safely use the safely use the out a full risk assessment
with minimal
equipment equipment. equipment. and minimise risks.
prompting.
and materials
 Students  Students must
 Students record  Students record choose the most effective
Makes and
record colorimeter colorimeter colorimeter way of recording
records
readings and plot readings and readings and plot measurements and
observations
calibration curve plot calibration calibration curve producing calibration
curve curve.
 Students  Students  Students  Students must
compare results compare results compare results research alternatives in
with normal with normal with normal order to plan their work.
Researches,
glucose glucose glucose Reporting covers the
references
concentrations concentrations concentrations planning, carrying out and
and reports
and identify and identify and identify an analysis of their results
reasons for reasons for reasons for in relation to normal
differences. differences.. differences. glucose concentrations.
: Very good opportunity : Good opportunity : Slight opportunity : No
opportunity
 A-level Biology required practical No. 11

Student Sheet

Production of a dilution series of a glucose solution and use of colorimetric

techniques to produce a calibration curve with which to identify the concentration of
glucose in an unknown ‘urine’ sample

Sugar in the urine is one of the first indications of diabetes.

Method

You are provided with the following:

 10 mmol dm-3 glucose standard.

 distilled water
 urine samples from Tom, Dick and Harry
 Benedict’s solution
 graduated pipettes (2 and 1 cm3) and pipette filler
 test tubes and test-tube rack
 water bath set at 90 oC
 colorimeter and cuvettes.

Prepare urine samples for testing

1. Label the test tubes with the name of the patient and add 2 cm3 urine samples from each
patient.
2. To each test tube add 2 cm3 Benedict’s solution. Mix the contents of the tube.

Prepare the glucose calibration curve

1. Label six test tubes 0 to 10 mmol dm-3 as shown in the table below.
2. Dilute the glucose standard (10 mmol dm-3) with water in the labelled test tubes and
complete the table to show volumes used to achieve each concentration.

Concentration of 0.0 2.0 4.0 6.0 8.0 10.0

final solution
/ mmol dm-3
Amount of water/ 2.0
cm3
Amount of glucose 0.0
standard /cm3

3. Add 2 cm3 of Benedict's solution to each tube. Mix the contents of each tube.
4. Place all the test tubes into the water bath together (including the tubes with the urine
samples) and time for four minutes. Allow to cool before taking readings from the
colorimeter.
5. Use the contents of the 0.0 mmol dm-3 glucose solution tube, which you have heated with
Benedict's, as a blank to calibrate the colorimeter to zero absorbance. Place the
remaining samples in cuvettes into the colorimeter and read the absorbance.
6. Record your results in a table and plot a graph of the absorbance of the known
concentrations of glucose.
7. Using the graph and the absorbance values obtained for the urine samples read off from
the graph the concentration of glucose in the urine samples.
8. Record your results in a suitable table.
A-level Biology required practical No. 11

Teachers’ Notes

Production of a dilution series of a glucose solution and use of colorimetric

techniques to produce a calibration curve with which to identify the concentration of
glucose in an unknown ‘urine’ sample

Materials

 glucose standard
 distilled water
 urine samples from Tom, Dick and Harry
 Benedicts solution
 graduated pipettes (2 and 1cm3) and pipette filler
 test tubes and test-tube rack
 water bath set at 90oC
 colorimeter and cuvettes.

Technical Information

Glucose standard solution 10 mmol dm-3

1.8 g glucose in 1 dm3 water

Urine samples (these can be varied)

Tom 0 mmol dm-3 glucose solution

Dick 5 mmol dm-3 glucose solution
Harry 8 mmol dm-3 glucose solution

Add weak tea to the water used to make the urine samples to colour the samples.

Benedict’s solution

Although results can be obtained using qualitative Benedicts solution, more reliable results
can be achieved using Quantitative Benedicts. CLEAPS Recipe Sheet 12 gives the
instructions for making up the quantitative chemical.

If the solutions are too dark when reacted with Benedict’s to use in the colorimeter, dilute the
glucose standard but still label it as 10 mmol dm-3

Risk assessment

Risk assessment and risk management are the responsibility of the centre.

Trialling

The practical should be trialled before use with students.

Additional notes
Eye protection should be worn should be worn when using Benedict’s solution.