Académique Documents
Professionnel Documents
Culture Documents
Catacutan,Kenneth
Leal,Catalina
Alambra, Karen
Date Submitted:
Experiment: CARBOHYDRATES
OBJECTIVES:
a. the objectives is to know whether a specific substances contains sugar or not using
Molisch’s test
Benedict’s test
c. To know whether a substance contains pentose sugar using Bial’s Orcinols test
Barfoed’s test.
CARBOHYDRATES
Theory
Carbohydrates include polyhydroxy aldehydes or ketones and their by-products. They make up
most of our diet and serve as the major source of energy .The play an important part in metabolic
processes by furnishing the carbon chain in for compound synthesis by living organisms.
Monosaccharides or simple sugars are highly soluble in water,less soluble in ethanol and insoluble
in ether .They cannot further hydrolyzes to simpler units. They are either aldoses and ketoses
depending on the functional group present .They may also be classified into tetroses, pentoses or
hexoses depending on the number of carbon atoms they possess. Free monosaccharides are all
reducing sugars. They also exhibit mutarotation, which means they can exists in 𝛼- and 𝛽- forms
maltose, which are abundant in germinain barley: sucrose, also known as cane sugar or beet sugar
: and lactose or milk sugar,which does not taste very sweet and is not fermented by yeast. Most
polysaccharides found in nature function either as structural units, (e.g . cellulose) or for storage
The molisch test is the general test for carbohydrates. The sugars are mixed with 𝛼-napthol. The
test tube is inclined and concentrated 𝐻2 𝑆𝑂4 is added along the side of the tube ,Causing the
formation of a lower layer of acid. The concentrated sulfuric cid will dehydrate the sugar allowing
it to react with the alcohol forming furfural or hydroxymethyl furfural. Formation of a purple ring
at the interface of the two liquids will indicate the presence of a carbohydrate.
Bial’s test is used to determine the presence of pentoses and nucleotides that contain pentose
sugars. When pentoses are treated with orcinol, furfural are formed and they will yield a blue green
compound in the presence of ferric ions. The reaction is not specific for pentoses because other
compounds like trioses,uronic acids, and a certainn heptoses will also give blue or green products.
3. Benedict’s test
Benedict’s test is a very sensitive test done under mildly alkaline conditions. The reagent contains
Cu𝑆𝑂 4 , 𝑁𝑎2 𝐶𝑂3 and sodium citrate. The formation of a brick red precipitate of 𝐶𝑢2 𝑂 is considered
positive. Most aldehydes have the ability to reduce Benedict’s reagent.Other compounds like
formic acid, hydrazobenzene, phenols, phenylhydrazine, pyrogallol, and uric acid will also give a
4. Barfoed’s test
Barfoed’s reagent contains cupric acetate in dilute acetic acid and is uded to distinguish between
monosaccharides but not oligosaccharides .Disaccharides are less easily oxidized but are oxidized
if they undergo prolonged heating, causing hydrolysis of the disaccharides into monsaccharides
which will then give a positive result. The concentration of the sugar solutions used in this test
should be approximately the same, because the use of a more concentrated disaccharide solution
may give a faster reaction than that of a relatively more dilute monosaccharide solution may give
a faster reaction than of that of relatively more dilute monosaccharide solution. Unlike Benedict’s
test, Barfoed’s test is carried out under acidic rather than basic medium5. Seliwanoff’s Test
Ths test is used to differentiate ketohexoses from aldohexoses. Ketosehexoses react faster with the
solution containing hydrochloric acid and resorcinol than aldohexoses. The dehydrated
ketohexoses fom a bright cherry red condensation product, whie the aldohexoses yield only a
pale pink coloration,a negative results. In this test, prolonged heating of samples should be
avoided.
6. Tollen’s test
Sugar with aldehyde groups are capable of reducing Tollen’s reagent (an ammonical solution of
𝐴𝑔+ ) to form a gray to black precipitate. If the rection vessel is clean and rate of deposition is slow
MATERIALS
250 ml beaker
toluene)
dilute HCl
carbonate
800 ml of water
17.3 𝐶𝑢𝑆𝑂4
PROCEDURES
1. Molisch’s test -Knowing if the substances wine and soda water contains sugar. Sugar solution
a) Mix 4 ml of sugar solution and 2 drops of the Molisch reagent in a test tube. This
reagent and mix the contents by gently shaking the test tube.
c) Incline the test tube and cautiously add about 5 ml of concentration sulfuric
acid,allowing the acid to run down the side of the tube. Sulfuric acid is denser than
water and will form a lower layer. Note the color of the ring formed at the junction
d) In the same manner of adding acid, add sulfuric acid to the control tube. What do
you observe
e) Repeat the above test with 3% sample solutions on the other samples.
Wine Positive
Result:
SUMMARY OF RESULTS:
For the summary of results, using distilled water and sugar solution as a control, the wine
and produced a purple ring during the experiment which indicates a positive result.
CONCLUSION:
We conclude that the wine and soda water that is said to be contains zero carbohydrates still
contains sugar. The sugar on the samples get dehydrated by the sulfuric acid to form furfural
products. Those products reacts with sulphonated a-naphthol to give a purple colored complex.
Moderate:Orange.red
173 g sodium citrate and 100 g of anhydrous sodium carbonate in 800 ml of water with the
aid if heat. Filter if necessary. Dissolve 17.3 𝐶𝑢𝑆𝑂4 in 100 ml water, then add to the citrate
c) Observe the color the solution and note whether a precipitate was formed. A change in
Result:
Beer Moderate
Yakult Moderate
sugar
Honey unknown
Before After
SUMMARY OF RESULTS:
For the summary of results, the hue of the solution will identifies reducing sugar. After boiling in
the bath sugar solution color stays unchanged presence of reducing sugar is absent. Yakult and
beer has a complexion yellow-orange that indicates the moderate concentration of reducing sugar
present in the solution. The energy drink has a brick red has a large amount of reducing sugar
while the flour has trace of reducing sugar as it turned into green.
CONCLUSION:
We conclude that Benedict’s reagent starts out blue. As it is heated in the presence of reducing
sugars, it turns yellow to orange. The “hotter” the final color of the reagent, the higher the
concentration of reducing sugar. In general, blue to blue green or yellow green is negative,
yelllowish to bright yellow is a moderate positive and bright orange is very strong positive.
3. Bial’s Orcinol Test- To know if the substance contains pentose sugar.
(NOTE: Bial’s Orcinol reagent dissolce 0.5 g of orcinol (3,5 dihydroxy toluene) in 250 ml
of 30% HCl, to which 10- 15 drop of 10% 𝐹𝑒𝐶𝑙3solution has been added.
c. Carefully heat each over a Bunsen flame until the solution begins to boil. Add 1-2 drops of
Result:
Beer Negative
Yakult Negative
Flour Negative
Honey Negative
Sucrose Negative
Before :
After:
SUMMARY OF RESULTS:
For the summary of results, the yakult,beer,energy drink,flour and honey didn’t produced a
blue-colored product that indicates a negative result while xylitol solution produced a blue-colored
CONCLUSION:
We conclude that arabinose contains pentose sugar, a five carbon sugar. Arabinose form
furfural in acidic medium which condense with orcinol in presence of ferric ion to give blue green
colored complex.
b. Add 4 ml of Seliwanoff’s reagent to each test tube.( NOTE: Seliwanoff’s reagent dissolve
11.5 g resorcinol in a liter of dilute HCl( 1 part concentration HCL to 2 parts of water )
c. Place the tube in water filled with boiling water and allow them to stay there for exactly 1
minute.
d. Note the changes and record which test tube gives a positive results in the shortest time.
Record the time required for a positive test for each sample
Substance tested Results
Beer Negative
Yakult Positive
Flour Negative
Honey Negative
Before After
SUMMARY OF RESULTS:
For the summary of results, the sucrose, yakult and energy drink change into a red complexion
that indicates the ketose sugar is present in the solution while the flour , honey and beer formed other
color means it is a negative. The flour and honey has a hue of light pink indicates the solution contains
aldoses sugar.
CONCLUSION:
We conclude that Seliwanoff ’s reagent when added to a solution containing ketoses a red color
is formed rapidly indicating a positive test. When added to a solution containing aldoses, a slower forming
light pink is observed instead. This test relies on the principles that, when heated ketoses are more rapidly
disaccharides
b. Add 3 ml of Barfoed’s reagent in each test tube. (NOTE: Prepare Barfoed’ reagent by
dissolving 60.5 g cupric acetate crystals in enough distilled water to make 1 liter. Filter and
d. Place all the tubes in a boiling water bath for 10 minutes. Record your observations.
Result:
Sugar Solution
Beer Disaccharide
Yakult Disaccharide
Flour Disaccharide
Honey Monosaccharide
SUMMARY OF RESULTS:
For the summary of results, the beer, yakult and flour has a blue complexion identifies
disaccharides while the honey has a brick red color that indicates the presence of monosaccharides.
CONCLUSION:
We conclude when Barfoed’s reagent ix with the solutions and heated in boiling water
bath, they react and crystal precipitate is formed. Copper acetate which is present in the barfoed
reagent convert to copper oxide and give brick red precipitate when react with monosaccharide.
6. Tollen’s test- To know whether a substance contains an aldehyde groups using Tollen’s test.
b) Add 2 ml of Tollen’s reagent into each tube. (Note: Prepare Tollen’s reagent by adding 1
𝑁𝐻4 𝑂𝐻+ 5ml of water) until the brown precipitate of silver oxide that form just dissolves.
This reagent must be prepared fresh and not sored since it decomposes when left standing
and yields and explosive decomposition product. Distract all left- over materials
Yakult Positive
Flour Negative
Honey Positive
Before After
SUMMARY OF RESULTS:
For the summary of results. The sugar solution, honey and flour change into a gray
complexion that indicates the a positive test while the flour, energy drink and beer formed other
CONCLUSION:
We conclude that sugar solution , honey and flour turns into gray because it has a presence
of aldehyde, aromatic aldehyde, and alpha-hydroxy ketone functional groups .Tollen’s reagent into
metallic silver which if the test is carried out into a clean glass test tube,forms a mirror on the test