Lab 04: Determination of the content of reducing sugar using Benedicts test

2007 Al-Bio Marking Scheme

Lab 4: Determination of the content of reducing sugar using Benedicts test

[Tested sample : Lucozade (glucose content : 22.48 %]

Area B1 (max. 15 marks)

Aim: ( max. 1 )
- By use of Benedicts test to estimate the concentration of reducing sugars
- in the given sample, Lucozade
- in term of glucose equivalent
- by comparing its reducing power with the standard glucose solution (2%)
Principle: (max. 6)
(Experimental design and principles)
Benedicts test
- condition / test
boiling of Benedicts solution with reducing sugars / test with Benedicts solution will give brick red
precipitate
-

mechanism
due to presence of blue solution Cu 2+ in Benedicts solution changed into insoluble brick-red precipitate
(Cu+)

observation and indicator for estimation

making the resulted mixture have a gradual change of colour with amount of reducing sugars inside the
solution : from blue to green, then yellow and finally brick red
the colour of boiled mixture / the amount of precipitate at bottom of test tube as indicator of comparing
concentration of reducing sugars.
the more the ppt. more reducing sugar

Expected result
similar ppt. between sample (may be diluted) and certain concentration of standard solution similar conc.
of reducing sugar

Variables:
independent variable: conc. of reducing sugars in solutions
dependent variable: amount of precipitate formed
Others:
pre-experimental trial:
to determine which of the solution (Lucozade or 2% standard glucose solution) with a higher conc. for
dilution
provide a range of solution in different conc. that include the conc. of the other solution.
assumption:
Page 1/4

Lab 04: Determination of the content of reducing sugar using Benedicts test

2007 Al-Bio Marking Scheme

1. the only reducing agent for precipitate formation in the experiment is reducing sugar in the solutions
2. the gas bubbles contained in Lucozade do not affect the measurement of solution volume.
Others

List of apparatus

Page 2/4

Lab 04: Determination of the content of reducing sugar using Benedicts test

2007 Al-Bio Marking Scheme

Procedures (max. 7)
Steps
Labelling
serial dilution

Benedicts test
- Benedicts solution added
- Initial observations
- Boiled in water bath

Quantities and Units

Volume with unit:
water, standard glucose and sample
resultant concentrations
-

Final observations
- place the boiled tube on test tube rack for:
compare colour differences immediately; or
wait for cooling and settlement of precipitate and compare amount of ppt.

volume of Benedicts solution

time period

Time waited

Precautions / control

Explaination

Pipette should be erect and be read at the same

level as eyes

Prevent reading errors

separate pipette for G , L and water transfer; or

wash pipette throroughly before pipetting
different fluids.

Prevent contamination of solutions with each other

excess Benedicts solution used

For complete reaction of all reducing sugars

heating in water bath

For even heating

sufficient heating duration

For complete reaction between reducing sugars and copper

(II) ion

water boiled with Benedicts solution (i.e.

control)

To prove no contamination in water and reagents / apparatus.

Others (max. 1)
- technique descriptions, good table presentation, others

Page 3/4

Lab 04: Determination of the content of reducing sugar using Benedicts test

2007 Al-Bio Marking Scheme

Area B2 (max. 8)
- Results recorded :
-- initial observation --- color of solutions (Lucozade + Benedicts solution greenish in colour)
--- any ppt.
-- final observation and measurements

--- color of solutions / supernatant)

--- amount of ppt.

Data table:
-- correct and proper title for all tables
-- proper heading (tube labels, tube contents conc. of solution, observation, height of brick red ppt. ) with
S.I. unit (if needed)
-- formula for calculation of dilution concentration (if not be mentioned in procedure)
-- complete table lines

Graph:
-- correct and proper title for all and graphs
-- correct x-axis (conc. of reducing sugar in term of glucose equivalent) and y-axis (amount of brick red ppt.
Formed) with proper unit and scale
-- best fit line of standard curve passing through origin (may be extended for estimation of unknown)

Correct comparison of ppt. content / pointer in graph showing unknown conc.

Estimation of original reducing sugar conc. of Lucozade with calculate (answer in term of glucose
equivalent)

Area B3 (max. 7)
- data description and interpretation
- other significant discussion (e.g. abnormalities, limitation, sources of error, assumption, improvement
suggestionetc.)
- conclusion: The original conc. of reducing sugar in Lucozade is __ % in term of glucose equivalent.
* with reference to the above guidelines, reports were marked by overall impression.

Page 4/4