Benedict

LEAL, MARK JUSTIN M.
BSN 1-A
A.Benedict’s Test
In order for us to check the presence of reducing sugar in a
sample, we need to perform a chemical test known as Benedict’s
Test. Before, to confirm if one has diabetes, they check the
presence of glucose in a urine. It also identifies reducing sugar
containing free ketone or aldehyde functional groups.
For the materials to prepare Benedict’s reagent [100mL],
we need Copper sulfate pentahydrate of about 1.73 gms, 10 gms
of Sodium carbonate, 17.3 gms of sodium citrate and distilled
water of up to 100mL.
To perform the Benedict’s Test, we need to mix 1mL of
sample to 2Ml of Benedict’s reagent. Second, we heat the
solution for 3.5 minutes. Lastly, we observe if there are changes
in the color.
For the interpretation, the blue color is our reagent color.
Depending on the percentage of reducing sugar in the sample,
the precipitate color is green if it has 0.5% of reducing sugar,
yellow if it has 1.0% of reducing sugar, orange if it has 1.5 % of
reducing sugar and red if it has 2% of reducing sugar.
To conclude, Benedict’s test is a test used to check the
presence of sugar in urine. We used the copper (II) sulfate as our
source of Cu2+ ions, the sodium carbonate as our alkaline
medium and the sodium citrate formed complexes with Cu2+
ions.
B. Benedict’s Reagent Preparation
Benedict’s Test is a biochemical test commonly employed
to detect the presence of reducing sugars with free aldehyde or
ketone functional groups, specifically hydroxy ketones.
Reducing sugars include almost all monosaccharides such as
galactose, glucose, glyceraldehyde, ribose, and xylose etc. and
some disaccharides like lactose and maltose.
To prepare Benedict’s Reagent, we need 1.73g of copper
sulphate pentahydrate, 10g of Anhydrous Sodium Carbonate,
17.3g of Sodium Citrate, 100ml of Distilled Water and 100 mL
volumetric flask. We can also purchase readymade Benedict’s
reagent solution but we will manually prepare it in the lab. For
the additional materials, we need two small beakers, two stirring
rods, small funnel, suitable container for storing the reagent, hot
plate or a burner, watch glasses and spatulas.
For the procedure of Benedict’s qualitative reagent, we will
weigh 1.73g of copper sulphate pentahydrate, 10g of Anhydrous
Sodium carbonate and 17.3g of Sodium citrate in separate watch
glasses using analytical weighing scale. Second, we will transfer
the sodium carbonate and the sodium citrate into a beaker.
Third, we will add of about 30-40 ml of distilled water and stir
the contents. In order to speed up the dissolving materials, we
will heat it. After which the contents have dissolved, we will
keep the beaker aside and transfer the copper sulphate into a
separate beaker. We will then add of about 10ml of distilled
water and stir to dissolve. Now, we are ready to add the copper
sulphate solution in small amounts into the beaker containing
the carbonate citrate solution while stirring the contents with
each addition. When the two solutions have been mixed, we will
rinse the copper sulphate beaker and the stirring rod using small
amounts of distilled water into the beaker containing the mixed
solution. Now, it’s time to transfer the solution into a 100ml
volumetric flask. Next is we will make up the final volume in
the volumetric flask to exactly 100ml with distilled water by
using a dropper or a pipette. Lastly, we will transfer the
contents of the volumetric flask into a suitable container with a
tight-fitting cap and label it as “Benedict’s Reagent”.
To sum up, Benedicts reagent is discovered by Stanley
Rossiter Benedict. It is a complex mixture of sodium carbonate,
sodium citrate and copper sulphate dissolved in distilled water.
Sodium carbonate provides the alkaline conditions which are
required for the redox reaction to occur between the reagent and
the reducing sugar contained in the test sample. Sodium citrate
acts as chelating agent and complexes with the cupric (copper
(II)) ions of copper sulphate so that they do not deteriorate to
cuprous (copper(I)) ions during storage.
C. Detection of sugar in urine
Benedict’s test is used to test for simple carbohydrates.
Benedict’s test identifies reducing sugars (monosaccharides and
some disaccharides), which have free ketone or aldehyde
functional groups. Benedict’s solution can be used to test for the
presence of glucose in urine. Benedict’s solution is a deep-blue
alkaline solution used to test for the presence of the aldehyde
functional group, – CHO.
In order for us to make 1 liter of Benedict’s solution we
will be needing 100 g of anhydrous sodium carbonate, 173 g of
sodium citrate and 17.3 g of copper (II) sulfate pentahydrate.
For the procedure of Benedict’s Test, we will place
approximately 1mL of the sample in a clean test tube and 2 ml
(10 drops) of Benedict’s reagent (CuSO4) in the test tube. The
solution is then heated in a boiling water bath for 3-5 minutes.
We will observe for color change in the solution of test tubes or
precipitate formation. If the color upon boiling is changed into
green, then there would be 0.1 to 0.5 percent sugar in solution. If
it changes color to yellow, then 0.5 to 1 percent sugar is present.
If it changes to orange, then it means that 1 to 1.5 percent sugar
is present. If color changes to red, then 1.5 to 2.0 percent sugar
is present. And if color changes to brick red, it means that more
than 2 percent sugar is present in solution.
The Benedict’s test is positive if we observe a formation of
a reddish precipitate within three minutes, there is the presence
of reducing sugars. Example of this is Glucose. On the other
hand, the test is negative if we observe that the color doesn’t
change and it remains blue. It is an example of Sucrose.
To sum up, Benedict’s quantitative reagent contains
potassium thiocyanate and is used to determine how much
reducing sugar is present. This solution forms a copper
thiocyanate precipitate which is white and can be used in a
titration. The titration should be repeated with 1% glucose
solution instead of the sample for calibration. The red copper(I)
oxide formed is insoluble in water and is precipitated out of
solution. This accounts for the precipitate formed. As the
concentration of reducing sugar increases, the nearer the final
color is to brick-red, and the greater the precipitate formed.
Sometimes a brick red solid, copper oxide, precipitates out of
the solution and collects at the bottom of the test tube.

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LEAL, MARK JUSTIN M.

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