6.4.7

Measuring Glucose Concentration

Test yourself

Colorimetry

Colorimetry is a technique used to identify the concentration of glucose in an unknown 'urine' sample. The steps involved are:

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1) Prepare urine samples

  • Take three test tubes and label each with the name of the patient (urine sample).
  • Add three cm3 urine samples from each patient.
  • To each test tube add 2 cm3 of Benedict’s reagent and mix the contents of the tube.
    • Benedict's reagent is used to detect the presence of glucose.
    • The reagent is blue and turns from blue to red when glucose concentration is greater. The solution also becomes more opaque.
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2) Prepare the calibration curve

  • Label six test tubes 0 to 10 mmol dm−3.
  • A glucose solution of 10 mmol dm−3 is provided. Dilute this standard glucose solution using distilled water to produce 5 additional concentrations which will be used to make the calibration curve.
  • The concentrations should be 0 mmol dm−3, 2mmol dm−3, 4mmol dm−3, 6mmol dm−3, 8mmol dm−3, and 10mmol dm−3.
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3) Incubate test tubes

  • Add 2 cm3 of Benedict's reagent to each tube and mix the contents.
  • 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.
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4) Use the colorimeter

  • 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.
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5) Record the results

  • Record your results in a table and plot a graph of the absorbance of the known concentrations of glucose. Plot concentration of glucose on the x-axis and absorbance on the y-axis.
  • 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.
  • Record your results in a suitable table.

Jump to other topics

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8The Control of Gene Expression (A2 only)

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