Nursing Biochemistry

Laboratory Report

Carbohydrates 1: Common
Reaction
Activity No. 3

Sanaani, Nur-Fatima M. (Principal Author)

Lui, Sharah Ghyll P.
Jasim, Nerizza B.
Locker No.33; NurBio Lab C
1st Semester, SY 2019-20
Nursing Biochemistry
Laboratory Report

RATIONALE
On this activity the distinction of carbohydrates are being test through various experiments that
involves: the common and specific reaction.

Carbohydrates are considered as one of the main bio-molecules and the most abundant
compounds that could be found in every food that people take. It makes up 80%.of the daily food that
people usually take. For it gives a primary fuel for a living organism to function. The word
“carbohydrate” is derived from the French word “hydrate de carbon” and is a term that is also a synonym
of “saccharide”, inclusive of sugars, starch and cellulose. Both of these terms are actually used
interchangeably when referring to them by modern scientific standards.

Figure .1 (Carbohydrates )

Polyhydroxyl aldehydes and ketones or substance that hydrolyzed to yield polyhydroxy aldehydes and
ketones, are composed of various carbon, oxygen and hydroxyl group. Aldehydes can be distinguish with
the presence of ( -CHO) and its carbon chain, while ketones are identified with the presence of (=O) that
are both a constitute the major groups in carbohydrates. Carbohydrates are mainly classified according to
how they are like in monosaccharide (1 saccharide), disaccharides (2 saccharides), and polysaccharides
(two to more saccharides).

Aldehydes and ketones are members of the same type of compound known as carbonyl
compounds. Carbonyl compounds are those that have carbon to oxygen double bonds attached.
In ketones the molecule is normally arranged so that the carbon to oxygen double bond can be the first
thing seen on the molecule

Aldehydes on the other hand have hydrogen attached to the carbon to oxygen double bond. It is this
hydrogen that accounts for the difference between aldehydes and ketones. it is important to note that this
hydrogen is what makes an aldehyde such a great reducing agent and thus can be oxidized easily. Ketones

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are however resistant to oxidation meaning they aren’t that easily oxidized. When an aldehyde is oxidized
in an acidic environment a carboxylic acid will be formed but when oxidized in an alkaline environment a
salt of a carboxylic acid will be formed.

Sugars are classified as reducing or non- reducing based on their ability to act as a reducing agent. A
reducing agent donate electrons during a redox reaction and is itself oxidized. The aldehyde functional
groups is the reducing agent in reducing sugar. Reducing sugar have either an aldehyde functional group
or have a ketones group, in an open chain form, which can be converted in to aldehyde. Reducing sugars
are simple sugar and includes all monosaccharides and most disaccharides. Exampls of the
monosachharides are glucose, fructose, and galactose, etc. example of reducing disaccharides are lactose,
and maltose.be reminded that disaccagrides have a one specific sugar that don’t reduce example ais
sucrose. In fact the sucrose solution is the most common non-reducing sugar.

Objective:

I. To identify some physical and chemical characteristics of typical carbohydrates

II. To Differentiate between monosaccharides, disaccharides and polysaccharides using the tests
included above

DATA INTERPRETATION:

Molisch’s test

This is common test for all carbohydrates. Carbohydrates are treated with concentrated sulfuric acid or
concentrated hydrolic acid. The compound of the acids will react to the -OH group of sugar are removed
in the form of water and furfural or violet degraded figure. These products reacts with sulphonated α-
naphthol to give a purple (violet red) colored complex. These products condensed with α-naphthol to
form purple condensation product.

(Lactose) (Glucose) (Sucrose) (Xylose) (Starch) (Unknown Sugar)

The experiments molisch’s test has a specific materials and substance that are used for examining which
composition of substance were they. Which basis do they belong Pentose and Hexose. On the test-tube 6

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it was labeled unknown sugar for it is still unknown, and need to be tested by various experiments to
determine its property. While on the other hand test tube 1 is labeled as Glucose, test tube 2 labeled as
Xylose, test tube 3 labeled as Lactose, test tube 4 as Sucrose, and test tube 5 labeled as Starch. While
adding a drop of molisch reagent there was a heat that is form within the mixed agents and that is being
transferred in the test tube. The after a while a 1ml of concentrated sulfuric acid H2 SO4 that is being
added to the mixed agent, making sure that it has being poured on the internal walls of the test tube. Not
long enough the reagent slowly formed a furfural formation also known as “violet ring conjunction or
violet ring”:

Figure .3( molisch ‘s structure, and equation)

Table No. 1

A. Molisch Test
Test Solutions Result Explanation

1% Glucose Violet This shows a positive result

ring because glucose is a
carbohydrate that has a several –
OH groups which the molisch
reagent would react.
1% Xylose Dark This shows a positive result
violet because xylose is a
full monosaccharide that has a
unifor several –OH groups which the
m molisch reagent would react.
1% Lactose Violet This shows a positive result
ring because lactose is a carbohydrate
that has a several –OH groups
which the molisch reagent would
react.

This shows a positive result

1% Sucrose Violet because sucrose is a carbohydrate
ring that has a several –OH groups
which the molisch reagent would
forma react.
tion

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Laboratory Report

1% Starch Violet This shows a positive result

ring because sucrose is a
forma polysaccharides that has a
tion several –OH groups which the
molisch reagent would react.

1% Furfural N/A

Unknown Sugar Purple ring The result of unknown sugar

formation shows a positive result because
theoretically the unknown sugar
is any sugar that can be either
monosaccharide, disaccharide, or
polysaccharide that are all under
carbohydrate that has a several –
OH groups which the molisch
reagent would react.

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DATA INTERPRETATION:

(theoretical results)Alkali:

The behavior of seven crystalline saccharine anilides on the paper chromatogram has been observed. The
R F values vary inversely with the molecular weight, and are greater for the α- Metasaccharinic anilides
than the β-isomers. The periodate oxidation of milligrams quantities of anilides and subsequent paper
chromatography of the fragments identify the “ metasaccharinic acids”. Compound that contains a metal
covalently bounded to a carbon atom of an organic species are called the “organometal compound”. The
properties and reactivities of organometallic compound differ greatly from those of either the metallic or
organic compound. These volatile , low-melting-point solid or liquid can be sublimed or distilled at
relatively low temperature and are solute in non-polra solvent.

Table No. 2

B. Action of Alkali (Search for Theoretical Result Only)

Alkali Test Result Explanation

Solution

10% NaOH 1%
Glucose

1%
Fructose

1%
Galactose

1% Ba(OH)2 1%
Glucose

1%
Fructose

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1%
Galactose

DATA INTERPRETATION: (THEORETICAL REPORT) *due to the lack of time*

Fehling’s test:

It is a test that indicates the reaction for reducing groups such as aldehyde and ketones functions. The
Fehling test is an indicator for aldehyde. Ketones cannot be oxidized using the Fehling reagent. However,
the Fehling test also reacts to ketoses. It makes it possible to differentiate between reducing and non-
reducing sugars solution using the blue alkaline cupric hydroxide solution, that is being heated with
reducing sugars it oxidize and turned into to yellow or red cuprous oxide and is precipitated. That leads to
the formation of the yellow or brownish-red precipitate helps that indicates positive results.

(Figure .5 Fehling’s)

Fehling's test can be used as a generic test for monosaccharides. It will give a positive result
for aldose monosaccharides (due to the oxidisable aldehyde group) but also for ketose monosaccharides,
as they are converted to aldoses by the base in the reagent, and then give a positive result. For this reason,
Fehling's reagent is sometimes referred to as a general test for monosaccharides.

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Laboratory Report

Benedict’s test:

Carbohydrates with a free aldehyde and ketones, groups in the reducing sugar. Reduce cupric hydroxide
in an alkaline with a medium red color cuprous oxide. Depending on the concentration of which sugar.
Yellow to green color reaction is a develop all monosaccharide’s which are distinguish a reducing sugar
as they all have a free reactive carbonyl group some. Disaccharide’s , like maltose, have exposed carbonyl
group are also reducing sugar, but less reactive than monosaccharide’s.

Figure.6( Benedict’s Equation)

In the laboratory, we used benedict substance, reagent to test out the particular reducing sugar of,
Glusoce, and other solution substance. Benedict reagent had begin to diffused into a aqua blue, by heating
up a maximum of 2 minutes water bathing the substance turned into reaction of yellow and orange” the
hotter” the final reaction of the substance of reagent. The higher the concentration of reducing sugar.

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Barfoard’s Test:

A barfoed’s is aa test where is to detect the presence of monosaccharides( reduction) sugar solution.
Barfoeds reagents, a mixture of ethanoic( acetic) acids and the formation of the copper II acetate, then
add in the substance of test solution which are glucose, and other sugar solution. Aldose and the kentose
can reduce cupric ion even in acidic condition. This test is used to distinguished a reducing the
monosaccharides from disaccharides. Monosaccharides react very fast where in disaccharides react very
slow.

A blue copperII oxide precipitate that formed will indicate the presence of the reduce sugar. The other
reaction would negative due to a presence of the disaccharides sugar because of their weak reducing
agents. The specific test is for monosaccharides, due to the weak acids of the nature of the barfoed
reagent. It is reduced only by monosaccharides.

Table No. 3

C. Reduction Test
*Within the table the Fehling’s test and results are all theoretical answer since the group was not
able to perform it*
Name of Test Test Result Explanation
Solution
1% There’s a red A non-reducing sugar
Fehling’s Test
glucose precipitates has because the carbon
formed within elements of the
the agent. aldehyde groups are
bonded
1% There’s a red A non-reducing sugar
xylose precipitates has because the carbon
formed within elements of the
the agent. aldehyde groups are
bonded
1% There’s a red A non-reducing sugar
fructose precipitates has because the carbon

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formed within elements of the

the agent. aldehyde groups are
bonded
1% There’s a red A non-reducing sugar
lactose precipitates has because the carbon
formed within elements of the
the agent. aldehyde groups are
bonded
1% There’s a red A non-reducing sugar
maltose precipitates has because the carbon
formed within elements of the
the agent. aldehyde groups are
bonded
1% There’s a red Sucrose is A non-
sucrose precipitates has reducing
formed within sugar because the
the agent. carbon elements of the
aldehyde groups are
bonded in what's
called A glycosidic
bond
1% N\A
starch
1% N\A
glycogen
Unknown N\A
sugar
1% Yellowish Low amount of the
Benedict’s
glucose reducing sugar present
Test
1% Dark Orange Moderate amount of
xylose reducing sugar present
1% Light orange Moderate amount of
fructose reducing sugar present
1% Dark Organge Moderate amount of
lactose reducing sugar present
1% Dark Orange Moderate amount of
sucrose reducing sugar present

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1% Light green Trace amount of

starch clear reducing sugar present
1% Light Blue Positives development
Barfoed’s Test
gluctose of the reagent because
of the classification of
the sugar solution of
monosaccharide’s
with in the 3-
2munites
1% Blue green Positives development
fructose Slightly light of the reagent because
of the classification of
the sugar solution of
monosaccharide’s
with in the 3-
2munites
1% Navy blue light Positives development
xylose of the reagent because
of the classification of
the sugar solution of
monosaccharide’s
with in the 3-
2munites
1% Light blue Negative reaction
lactose because of the
classification of the
sugar itself is a
disaccharides
1% skyblue Positives development
maltose of the reagent with in
the 3- 2munites
1% Aqua blue Negative reaction
sucrose because of the
classification of the
sugar itself is a
disaccharides
Unknown Light blue Positives development
sugar of the reagent with in

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the 3- 2munites

DATA INTERPRETATION:

Nylander’s :

In a chemical tests for the process and process of the redundancy of the detection of the
reducing of sugar solution, or oxidization of the sugar solution.In the case of alkaline, solution bismuth
sub nitrate. The bismuth hydroxide formed is reducesd to black metallic bismuth as shown in the concept
of equation,

Bi(OH)2𝑁𝑂3 + KOH Bi(OH)3 + KNO3

2Bi(OH)3( 𝑤𝑖𝑡ℎ 𝑟𝑒𝑑𝑢𝑐𝑖𝑛𝑔𝑓 𝑠𝑢𝑔𝑎𝑟 𝑎𝑛𝑑 ℎ𝑒𝑎𝑡) 2Bi+

In our experiment the solution that yield a positive result was Glucose, Maltose, Lactose ,And Fructose.
While Sucrose, Starch was negative indicate by a clear solution.

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Picric acid Test:

The picric aid test for carbohydrates is a very sensitive test, for the presence of the reducing sugar
reaction with the picric acids. The yellow crystalline solids also chemically known as 2,4,6-
trinitrophenol( TND) to form a red coloured picranic acids. All monosaccharides and disaccharides
containing the potentially a free aldehyde or ketone group possess reducing properties. The reduce came
from the organic acids when in the alkaline solution picrics acid ( yellow solution) to pircramic (
mahogany red solution). An appearance of red and color indication the presence of reducing sugar in the
solution.

Picric Acid Test: (Equation, and Structure)

Figure. 9

Tollen’s Test:

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Tollence test refers to the silver- mirror test, used for a distinguished between a aldehyde and
a ketones properties. It exploits the fact that aldehydes are readily of the process of which is oxidized
(seen oxidation) whereas the ketones are not. Tollen’s test uses a reagent known as tollen’s reagent, which
is a colorless reaction, which refers as (silver mirror), aqueous solution containing silver ion’s coordinates
to ammonia (Ag(NH3 )2+). Tollen’s reagent has oxidized an aldehyde into the corresponding carboxylic
acid.

Figure .12(tollen’s Test)

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Table No.4

Name of Test Test Result Explanation

Solution

Nylander’sn 1%glucose Black

Test precipitate

(Search for 1%fructose Olive green

Theoretical
Result Only)

1% xylose Black

1%lactose Light yellow

1% Dark yellow
maltose

Picric Acid 1% glucose Red orange Positive result for

Test picric acid test
indicates the
presence of
reducing sugar

1% Red colour Positive result for

fructose picric acid test
indicates the
presence of
reducing sugar

1% xylose Slight red Positive result for

picric acid test
indicates the
presence of
reducing sugar

1% lactose Orange Positive result for

colour picric acid test
indicates the
presence of

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reducing sugar

1%maltose Clear no Negative (non-

reaction reducing
presence, did not
change in a
Yellow, and
Mahogany red
solution.

Tollen’s Test 1% glucose Clear No Corresponds to

reaction the aldehydes
carboxylic acid.

( silver mirror)

Classification of unknown sugar Monosaccharides

ANSWERS TO QUESTIONS

1. Why do all sugars give a positive result for molisch’s test?

ANSWER: All catbohydrates whih are the monosaccharides, disaccharides, and th

polysaccharides give positive reaction for molisch test. Due to its based on its dehydration of
the carbohydrates by the sulphuric acids to produce a aldehyde, which condence the two
molecules of the α- naphthol, which result of the appearance of the substance, the purple ring
conjuction of the interface.
2. What compound other than α- naphthol can be used as a reagent for the test??

ANSWER: molisch test is a test for the presence or determination of carbohydrates in a

sugar solution by the use of molisch reagent or the alpha- napthol and concentrated sulfuric
acids (H2SO4, used preferably and often) event in our laboratory experiment which is the
molisch’s test.

3. Why is it important to use freshly prepared Fehling’s reagent?

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ANSWER:the solution have to be freshly prepared because fehlings reagent is used as an

oxidizing agent and if not freshly prepared it may not oxidize aldehyde to an acid as
Cu(OH)2 Gradually losse its reactivity as it reacts with oxygen.

4. Compare the action of a strong and a weak alkali on a reducing sugar?

ANWERS: alkalis and acids can be described as strong or weak. This does not mean the same
as concentrated or dilute. The strength of an alkali depends on how ionized it is in water. An
alkali forms hydroxide ions (OH − ions) in water, a strong alkali is completely (100%) ionsed,
an examples of a strong alkali is Sodium Hydroxide a Strong alkali has a pH of 14. A weak
alkai is only partly (less than100%) ionized, an examples of a weak alkali is Ammonia.

5. How are the conditions for barfoed’s test different from those for Benedict’s test?

ANSWER: the test are extremely similar. Barfoed’s test detects monosaccharides through a
reducing reaction that precipitates blue copper(I) oxide and the benedict’s reagent detects
reducing sugar by producing the difference precipitated (yellow and orange ). The only real
difference is that benedict’s produces a greater volume of precipitate, and the precipitate can
vary in colors depending on the amount of sugar present
6. Account for the black precipitate in Nylander’s test?

ANSWERS: the Nylander test is to determine glucose in urin, the black precipitate is metallic
bismuth. It is formed when glucose or fructose reduce the bismuth oxynitrate in an alkaline
state.

CONCLUSION

Carbohydrates are important biomolecules, in the sense that they are pretty abundant on the surface of the
planet. Having a wide variety of industrial applications as well as already naturally occurring applications,
the actual importance of carbohydrates is already quite obvious. Ranging from the glycogen that exists as
the energy storage within animals, to the cellulose that forms the integral structure of plants,
carbohydrates can definitely be seen as something that is akin to the basic make up of life as we know it.
This experimental procedure has aided in being able to understand the structures of these carbohydrates
and how these physical properties dictate how the sugar will behave in different situations and the like.
Like how sugars are considered to be reducing sugars if they have an aldehyde group that can be oxidized
or if they are able to create one through isomerism (tautomeric conversion of fructose to an aldose).
Moreover, the knowledge of the fundamental classification test for these compounds has been invaluable,
given that the knowledge and familiarity with these tests also reinforce the knowledge of the properties of
the carbohydrate samples tested.

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REFERENCES
http://www.namrata.co/category/practical-biochemistry/reactions-of-carbohydrate
• "Laboratory Tests for Cabohydrates"
http://www.slideshare.net/katealyssacaton/seliwanoff-benedicts-test
• "Testing for Cabohydrates"
https://biokamikazi.files.wordpress.com/2013/10/merged_document.pdf
• "Benedict's Test"
www.harpercollege.edu/tm-ps/chm/100/dgodambe/thedisk/carbo/bened/benedict.htm
• "Drawing Sugar Structures"
www.chtf.stuba.sk/~szolcsanyi/education/files/Chemia%20heterocyklickych%20zlucenin/Pred
naska%206/Odporucane%20studijne%20materialy/Drawing%20sugar%20structures.pdf
• "Carbohydrates"
chemed.chem.purdue.edu/genchem/topicreview/bp/1biochem/carbo5.html
• "Amylose"
en.m.wikipedia.org/wiki/Amylose
• "Benedict's Test"
www.harpercollege.edu/tm-ps/chm/100/dgodambe/thedisk/carbo/bened/benedict.htm
• "Benedict's Reagent"
www2.sunysuffolk.edu/sabatil/by14/by14lab3/benedict.htm
• "Reducing Sugars"
en.m.wikipedia.org/wiki/Reducing_sugar
• "Reducing Sugar"
www.ausetute.com.au/redsugar.html
• "Barfoed's Test"
www.harpercollege.edu/tm-ps/chm/100/dgodambe/thedisk/carbo/barf/barfoed.htm
• "Functional Tests for Cabohydrates"
https://www.msu.edu/course/lbs/145/luckie/Lab1.html
generalchemistrylab.blogspot.in/2011/12/iodine-test-for-starch-and-glycogen.html?m=1

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