EXPERIMENT NO.

GAS BURNER

GROUP: 1 INTRODUCTION AND OBJECTIVES (15%)

MATERIALS AND METHOD (15%)
NAMES: ACOBA, ALESSANDRA RIAN DATA ANALYSIS AND DISCUSSION (25%)
CASTRO, PRECIOUS NICOLE CONCLUSION (25%)
DE GRACIA, APRIL JAN HONEY DREW LITERATURE CITED (10%)
RAGUINDIN, JOHN RYAN REPORT FORMAT AND QUALITY (10%)
RAMORAN, ALTHEA
REYNON, KRYSHELLE
SIMISIM, RIZZA MAE TOTAL
TAYAMEN, DION DOMINIC

INTRODUCTION:

Carbohydrates are one of the three macronutrients in the human diet, along with protein and fat. They are a group of
naturally occuring carbonyl compounds, either an aldehyde or ketone group, as well as many hydroxyl groups. They are often
called polyhydroxy aldehydes or ketones. One of the functions of carbohydrates is to provide energy for the body by the breakdown
of foods that we eat. The energy that can be obtained is used for reactions that are necessary in the body (Holesh et. al., 2023).

Carbohydrates are also known as "saccharides," and are considered the most abundant organic molecules found in
nature. A monosaccharide consists of a single carbohydrate unit such as glucose and fructose. They cannot be further hydrolyzed
thus, called as simple sugars (Aryal, 2023). Sucrose and lactose and disaccharides and they contain two monosaccharides. Lastly,
polysaccharides consist of many monosaccharides that are linked together. Starch, pectin, glycogen, and cellulose are examples of
it.

Furthermore, most monosaccharides are in cyclic forms when it is in a solution. The aldehyde or ketone group reacts with
one of the –OH groups on the other end of the same molecule to form a cyclic hemiacetal. A furanose can be seen in fructose
where it is a 5-membered ring structure. On the other hand, a 6-membered structure is called pyranose and is found on glucose
molecules.

In connection with the above, two chemical reactions, namely Furfural-forming property and reducing property, are used in
detecting carbohydrates in a solution. Under the Furfural-forming property, carbohydrates undergo the process of dehydration with
the aid of non-oxidizing acids. This process will enable aldehydes to condense with aromatic amines and phenols thus, forming a
furfural or hydroxy furfural. The furfural formation can be used as a qualitative or quantitative test for carbohydrates. Moreover, the
reducing property as mentioned, the ring structures open into an open-chain structure that produces a free form of aldehyde or
ketone. Aldose sugar has a reducing property whereby precipitated products are formed such as cuprous oxide (Cu2O) by the
reducing agent cupric ion (Cu2+). In the case of ketose sugar, fructose for instance, undergoes tautomerism in which it can change
and generate glucose and mannose in alkaline condition. Heating the sample with an alkaline reagent which contains cupric ions is
the most common test used for detecting reducing sugars. Potential aldehyde or ketone groups are oxidized and the copper is
reduced to red cuprous oxide.

There are several chemical tests in identifying the presence of carbohydrates in a given solution. They can be classified
as confirmatory tests for aldoses and ketoses (Molisch test, Seliwanoff’s test and Moore’s test), and reduction tests (Fehling’s test,
Benedict’s test, and Barfoed test).

According to Aryal (2022), the Molisch test is used to detect carbohydrates in different samples. It detects the formation of
carbohydrates as a by-product and distinguishes them from other biomolecules. The presence of a blue or purple ring indicates that
there is a presence of carbohydrates. The absence of color indicates otherwise. The test uses a a-naphthol reagent and shows
positive for all carbohydrates. Monosaccharides give a rapid positive test while reactions are slower in disaccharides and
polysaccharides.

Likewise, the Seliwanoff’s test uses resorcinol and gives similar reactions. It is a test that distinguishes keto sugars from
aldo sugars. To indicate a positive result, a red color is formed depicting the presence of ketoses.

The Moore’s test is used to detect the presence of reducing sugars. As the reducing sugars are heated with sodium
hydroxide (NaOH), a color of brown is released indicating a positive result except for sucrose such that it lacks a free aldehyde
group making it a non-reducing sugar (Bharath, n.d.).

Moving onto the reduction tests, Fehling’s test aims to differentiate between aldehyde and ketone groups in a solution.
The identification of reduction sugar is done by the Fehling’s reagent which is composed of two solutions. For the Fehling reagent
test, sugars such as glucose, fructose, and lactose yield positive results (Singla, n.d.).
 Benedict's test checks the presence of a reducing sugar in a given analyte. By the use of this test, simple carbohydrates
that contain free ketone or aldehyde functional groups are identified. The formation of brick red precipitate indicates a positive
result for this test. Also, this solution has been used in clinical laboratories for testing urine (BYJU’S, 2021).

Lastly, the Barfoed’s test detects reducing monosaccharides. Under the condition of acidity or prolonged boiling,
disaccharides may yield a positive result. It is also different from Fehling’s and Benedict’s test in a sense that reactions occur in an
acid solution rather than in alkaline media. Also, the presence of chlorides makes it unsuitable for the detection of urine.

OBJECTIVE:

The experiment aims to enhance understanding of carbohydrates by investigating their molecular structure, reactivity, and
physical properties. It seeks to:

1. Distinguish between monosaccharides and disaccharides based on their molecular structure.

2. Understand how the structure of carbohydrates influences their solubility properties.
3. Predict the reactivity of various carbohydrates when reacting with different oxidizing agents.
4. Learn the structural differences between aldose and ketose sugars.

MATERIALS:

Laboratory Equipment Laboratory Glassware Reagent

 Wash Bottle  Test Tubes  Glucose
 Weighing Balance  5 mL pipette  Galactose
 Heating Set-up  500 mL beaker Fructose
 Rubber Aspirator Lactose
Sucrose
10% Sodium Chloride (NaCl)
0.2% Hydrochloric acid (HCl)
95% Ethyl alcohol
10% Copper Sulfate (CuSO4)
Concentrated Sulfuric acid (H2SO4)
6M Sodium Hydroxide (NaOH)
Molisch reagent
Seliwanoff’s reagent
Benedict’s reagent
Fehling’s Solution A and B
10% Glucose
10% Fructose
10% Sucrose
Barfoed’s reagent
METHOD:

EXPERIMENTAL PROCEDURE:
A. Macroscopic Appearance

Prepare ample Place each in a separate glass and

amount of glucose, using gloves, spread out the
galactose, fructose, sample to easily identify its form
lactose, and sucrose. and color.

Observe each sample for their

Prepare laboratory form and color (macroscopic
equipment such as appearance).
glass and test tube.

Record observations
in Table 1 on page 34
using powdered or
Take small amounts of crystallized for form,
powdered glucose, galactose, and white or colorless
fructose, lactose, and sucrose. for color.

Clean the glasses used in the

experiment, as well as the
laboratory area.
 B. Solubility

Prepare five sets of four test

tubes each for glucose,
galactose, fructose, lactose, and
sucrose: label them
accordingly.

Place 1 ml of water to each test

tube.

Add a tiny pinch of each Record observations

respective samples to the test on the samples’
tube. solubility in water.

In another set of labelled test

tubes, place 1 mL of 10% NaCl
solution to each tube.

Record observations
Add a tiny pinch of each on the samples’
respective samples to the test solubility in 10%
tube. NaCl solution.

For the third set, place 1 mL of

0.2% HCl to each tube.

Record observations
Add a tiny pinch of each on the samples’
respective samples to the test solubility in 0.2%
tube. HCl.

For the last set, place 1 mL of

95% ethyl alcohol to each tube.

Record observations
Add a tiny pinch of each on the samples’
respective samples to the test solubility in 95%
tube. ethyl alcohol.
 Record observations
and tabulate results in
Table 2 on page 34.

Dispose the mixtures, wash the

test tubes, and clean the area.
C. Confirmatory Tests for Aldoses and Ketoses
1. Moore’s Test (influence of Concentrated Alkali)
2.
Prepare five test tubes
for the same sample, a
Remove the test tubes from the
container to be used for
heated container.
heating, and a burner.

Mix 1 mL of 10% carbohydrate Record observations

solution to each sample and add on color changes in
1 mL of 6M NaOH. Table 3 on page 35.

Put the test tubes inside the Dispose the mixtures properly,
container used for heating and wash used containers and test
heat them in a water bath. tubes, store the burner, and
clean the area.

Wait for a few minutes until

another color gets produced
during the heating process.
 3. Molisch’s Test
Prepare five test tubes
for the same sample
(glucose, galactose,
sucrose, fructose, and Repeat the test with the other
lactose). carbohydrate samples.

Place 1 mL of 10%
Observe the color
carbohydrate solution (glucose,
produced at the
galactose, fructose, sucrose, and
interface of the two
lactose) in a test tube.
liquids and record
observations in Table
3.

Add 1 drop of freshly prepared

Molisch reagent and mix
thoroughly.
Dispose chemicals and mixtures
with care, wash test tubes, and
clean the area.
Incline the tube and allow 1 mL
of concentrated H₂SO₄ to flow
along the inner side of the tube
and forms a layer at the bottom
part of the tube.

Do not shake the tube. Wait

for a few minutes until a
color is observed inside the
tube.
 4. Seliwanoff’s Test
Prepare five test tubes
for the same sample, a
Remove the test tubes from the
heating container, and
heated container.
a burner.

Record the changes

To 1 mL of Seliwanoff’s and results in Table
reagent, add 5 drops of 10% 3.
carbohydrate solution (glucose,
galactose, fructose, sucrose,
and lactose).

Dispose the mixtures properly,

wash used containers and test
Put the test tubes inside the tubes, store the burner, and
container used for heating and clean the area.
heat them in a water bath.

Wait for a few minutes until

an appreciable changed is
noticed during the heating
process.
C. Reduction Test
1. Fehling’s Test

Prepare test tubes

for glucose,
fructose, lactose,
and sucrose.
Remove the test tubes from the
heated container.

Prepare a mixture of
Fehling’s reagent: A Record the color
and B. changes and results
in Table 4. If it
changes, it’s positive,
but if the color
remains, it’s
negative.
Mix the Fehling’s reagent to
each test solutions (1 mL of
10% carbohydrate solution)

Dispose the mixtures properly,

wash used containers and test
tubes, store the burner, and
Put the test tubes inside a clean the area.
container used for heating and
heat them in a boiling water.

Wait for 6-8 minutes and

observe the color changes.
2. Benedict’s Test

Prepare test tubes

for the same
samples (glucose,
fructose, lactose,
and sucrose)
Remove the test tubes from the
heated container to cool it down.

Prepare a Benedict’s
Record the color
reagent.
changes and results
in Table 4. If it
changes, it’s positive,
but if the blue color
of the solution
remains, it’s
Add 1mL of Benedict’s reagent negative.
to each of the 1mL of 10%
carbohydrate solutions and mix
the solutions thoroughly.
Dispose the mixtures properly,
wash used containers and test
tubes, store the burner, and
clean the area.

Place the test tubes in a boiling

water bath.

Wait for 6-8 minutes and

observe the color change of
each test tubes.
3. Barfoed’s Test

Prepare test tubes

for glucose,
fructose, lactose,
and sucrose.

Remove the test tubes from the

heated container.

Prepare Barfoed’s
reagent (copper
acetate in acetic
acid)
Allow the solutions to cool
down for 15 minutes and
observe the precipitate.

Add 1mL of Barfoed’s reagent

to each of the 0.5 mL of 10%
carbohydrate solutions and mix.

Record the color

changes and results
in Table 4.
Place the test tubes in a boiling
water bath.

Dispose the mixtures properly,

wash used containers and test
tubes, store the burner, and
Wait for exactly 1 minute for clean the area.
the solutions.
DATA and RESULTS:

Table 1: Macroscopic Appearance of Carbohydrates

OBSERVATIONS
CARBOHYDRATES
Form Color

GLUCOSE fine powder white

GALACTOSE fine powder white

FRUCTOSE crystalline granules colorless

LACTOSE fine powder white

SUCROSE crystalline granules colorless

Table 2: Solubility of Carbohydrates in Different Solvents

OBSERVATIONS

CARBOHYDRATES WATER 10% NaCl 0.2% HCI 95% Ethanol

GLUCOSE Soluble Soluble Soluble Insoluble

GALACTOSE Soluble Soluble Soluble Insoluble

FRUCTOSE Partially soluble Soluble Soluble Insoluble

LACTOSE Partially soluble Partially soluble Partially soluble Insoluble

SUCROSE Soluble Soluble Soluble Insoluble

Table 3: Confirmatory Tests for Aldoses and Ketoses

OBSERVATIONS

CARBOHYDRATES MOORE’S TEST MOLISCH’S TEST SELIWANOFF’S TEST

GLUCOSE Brown Purple ring No change

FRUCTOSE Brown Indigo ring Light orange

SUCROSE No change Indigo ring Light orange

LACTOSE Brown Purple ring No change

GALACTOSE Dark-brown Purple ring No change

Table 4: Reduction Tests for Carbohydrates

OBSERVATIONS

CARBOHYDRATES FEHLING’S TEST BENEDICT’S TEST BARFOED’S TEST

GLUCOSE Dark-brown precipitate Dark-red Red precipitate

FRUCTOSE Brown precipitate Dark-red Red precipitate

SUCROSE Brown precipitate Blue No change

 LACTOSE Dark-red precipitate Red-orange No change

DISCUSSION:

This section presents the observations made during the experiment, which focused on examining the macroscopic
characteristics of carbohydrates, specifically their form and color. The experiment also determined the solubility of carbohydrates in
various solvents, including water, 10% NaCl, 0.2% HCl, and 95% ethanol. Additionally, confirmatory tests for aldoses and ketoses
were conducted using Moore’s Test, Molisch’s Test, and Seliwanoff’s Test. Finally, reduction tests for carbohydrates were
performed utilizing Fehling’s Test, Benedict’s Test, and Barfoed’s Test to assess the reducing properties of the carbohydrates.

With varying observations, table 1 shows the physical characteristics of the carbohydrates—glucose, galactose, fructose,
lactose, and sucrose. Glucose, galactose, and lactose are characterized by their fine powder form, whereas fructose and sucrose
exhibit a crystalline, granular structure. All the carbohydrates consistently display a white coloration, highlighting their uniformity in
appearance, except for fructose and sucrose which displayed a colorless color.

Table 2: Solubility of Carbohydrates in Different Solvents, summarizes the solubility results of six carbohydrates when
mixed with various solvents. In water, glucose, galactose, and sucrose were observed to be soluble, while fructose and lactose
demonstrated reduced solubility. According to the study of Cardoso et al. (2012), the replacement of water by heavy water (D2O)
decreased the solubility of carbohydrates, indicating that they are more soluble in water. With 10% NaCl as the solvent, glucose,
galactose, fructose, and sucrose were soluble, whereas lactose remained less soluble. Similarly, when 0.2% HCl was used,
glucose, galactose, fructose, and sucrose were soluble, with lactose again being the exception. Finally, in 95% ethanol, all
carbohydrates exhibited reduced solubility, indicating uniform insolubility in this solvent.

Table 3: Confirmatory Test for Aldoses and Ketoses presents the results of three biochemical tests, namely, Moore’s Test,
Molisch’s Test, and Seliwanoff’s Test, which was performed to classify the carbohydrates as reducing sugars, aldoses, or ketoses.

In Moore’s Test, glucose, fructose, lactose, and galactose exhibited a color change from colorless to brown when heated
with sodium hydroxide (NaOH), confirming their classification as reducing sugars. The intensity of the coloration varied, with
galactose displaying the darkest hue. In contrast, sucrose did not undergo any color change, affirming its classification as a non-
reducing sugar. Calo (n.d.) to find out if there are reducing sugars present, utilize Moore's test. The smell of caramel is released
when reducing sugars are heated with sodium hydroxide (NaOH), turning yellow, orange, and finally dark brown.

In Molisch’s Test, all carbohydrates tested—glucose, fructose, sucrose, lactose, and galactose—yielded a positive result,
indicated by the formation of a purple ring. The purple rings of fructose and sucrose were lighter in comparison to the others, but
the test confirmed the presence of carbohydrates, distinguishing between monosaccharides (e.g., glucose, fructose, galactose) and
disaccharides (e.g., sucrose, lactose). Sapkota (2022b) a method for identifying the presence of carbohydrates in different samples
and differentiating them from other biomolecules is the Molisch test. It looks for the development of a purple ring at the interface
between the test solution and sulfuric acid; if there is no color, the test is considered a negative result.

Finally, in Seliwanoff’s Test, fructose and sucrose produced a light orange coloration, classifying them as ketoses.
Meanwhile, glucose, lactose, and galactose showed no significant color change, verifying their classification as aldoses. Sapkota
(2022a) the test is used to differentiate between sugars that contain ketone and aldehyde. Resorcinol and concentrated
hydrochloric acid are used as reagents. Ketoses produce a compound known as xanthenoid, which has a deep cherry red hue,
when they dehydrate more quickly than aldoses. If the test is extended, aldoses may react somewhat to create a pale pink to
cherry red tint. Other carbohydrates, such as sucrose and inulin, which are degraded by acid to produce fructose, are also detected
by the test. Ketoses are indicated by a positive result, whereas no ketoses are indicated by a negative result. Fructose in
fermentation media is determined colorimetrically using Seliwanoff's color reaction.

Table 4: Reduction Tests for Carbohydrates summarizes the results of reduction tests conducted on four different
carbohydrates using Fehling’s Test, Benedict’s Test, and Barfoed’s Test.

In Fehling’s Test, glucose exhibited a dark brown color without precipitate, indicating a negative result. Fructose and
sucrose produced a brown color with precipitate, signifying a positive result. Lactose showed a dark red color without precipitate,
yielding a negative test result. Fehling Test (n.d.) the test uses sodium hydroxide and copper sulphate solutions, with the latter
turning red when the reducing sugar reacts with the copper sulfate solution. The test involves heating the material, with a brick-red
precipitate indicating an aldehyde group and a reddish-brown precipitate indicating the presence of reducing sugars. A poor result
indicates a lack of reducing sugars.

In Benedict’s Test, glucose and fructose formed dark red precipitates, identifying them as moderate-reducing sugars.
Sucrose produced a blue precipitate, confirming a negative result, while lactose yielded a red-orange precipitate, also classifying it
as a moderate-reducing sugar. Aryal (2022) Benedict's reagent undergoes a reduction reaction when reducing sugars are added,
changing its color from green to brick or rusty-brown. The potassium thiocyanate-containing reagent produces a precipitate of
copper thiocyanate that can be utilized in titration. The solution turns orange or brick red as a result of the reaction, which happens
when the reducing property of simple carbohydrates is diminished. Benedict's test does not yield a positive result for complex
carbohydrates, such as starches, unless they are heated or digested.
 Lastly, in Barfoed’s Test, glucose and fructose formed red precipitates within minutes, confirming the presence of
monosaccharides. In contrast, sucrose and lactose showed no reaction, indicating them as disaccharides. Barfoed's test is a
chemical method used to identify monosaccharides by reducing cupric acetate to cuprous oxide, forming a brick-red precipitate.
Disaccharides react slower, and a red precipitate within two minutes indicates monosaccharides, while a precipitate after ten
minutes indicates disaccharides (Bhuyan, 2023).

CONCLUSION:

The experiment aimed to enhance the understanding of carbohydrates through the investigation of molecular
structure, reactivity, and physical properties. It also effectively met the objectives of observing the physical and chemical
properties of different carbohydrates through macroscopic appearance, solubility tests, and confirmatory tests and
reduction test. Glucose, Galactose, and Lactose have a fine powdery texture and also depicted a white color. On the other
hand, Fructose and Sucrose have a crystalline granules and are colorless. Furthermore, the solubility of the
carbohydrates is also tested. In water, Glucose, Galactose, and Sucrose are soluble, whereas Fructose and Lactose are
partially soluble. The solubility of carbohydrates in 10% NaCl showed that Glucose, Galactose, Fructose, and Sucrose are
soluble. Lactose was partially soluble. The use of 0.2% HCl also showed the same result where Glucose, Galactose,
Fructose, and Sucrose are soluble and Lactose is partially soluble. Lastly, all of the carbohydrates were insoluble in 95%
Ethanol.

Carbohydrates were also categorized using three biochemical tests: Moore, Molisch, and Seliwanoff tests.
Ketones such as Fructose and Sucrose were characterized as light orange, whereas Glucose, Lactose and Galactose
were classed as aldoses due to the no change under Seliwanoff’s test. Through the Molisch test, the presence of purple
ring in Glucose, Galactose and Lactose showed a positive result. The presence of fructose and sucrose were lighter in
comparison to others and confirms the presence of carbohydrates distinguishing glucose, fructose, galactose as
monosaccharides; while sucrose and lactose as disaccharides. For identifying the presence of carbohydrates, different
samples and other biomolecules is through the Molisch Test; with the development for a purple ring at the interference
between the test solution and sulfuric acid. In the Seliwanoff’s Test, fructose and sucrose produced a light orange
pigment classifying them as ketoses, while glucose, lactose, and galactose showed no change, they are classified as
aldoses. In Seliwanoff's test, fructose and sucrose produced a light orange color, identifying them as ketoses, while
glucose, lactose, and galactose showed no significant color change, confirming their classification as aldoses. This test
distinguishes between ketose and aldose sugars using resorcinol and concentrated hydrochloric acid as reagents.
Ketoses dehydrate faster than aldoses, forming a xanthenoid compound with a deep cherry-red color. If the test is
prolonged, aldoses may develop a pale pink to red hue. The test can also detect carbohydrates like sucrose and inulin,
which degrade into fructose under acidic conditions. A positive result indicates the presence of ketoses, while a negative
result indicates their absence. This colorimetric reaction is also used to measure fructose in fermentation media. The
reduction tests for carbohydrates involved Fehling's, Benedict's, and Barfoed's tests to analyze four different sugars. In
Fehling's test, glucose turned dark brown without forming a precipitate, indicating a negative result. Fructose and
sucrose formed a brown color with precipitate, signaling a positive outcome, while lactose showed a dark red color
without precipitate, also indicating a negative result. This test uses sodium hydroxide and copper sulfate, with a brick-red
precipitate indicating the presence of reducing sugars. A reddish-brown precipitate suggests a weaker reaction, while no
change indicates the absence of reducing sugars. In Benedict's test, glucose and fructose produced dark red precipitates,
classifying them as moderate-reducing sugars. Sucrose formed a blue precipitate, indicating a negative result, while
lactose yielded a red-orange precipitate, confirming a moderate-reducing sugar. Benedict's reagent undergoes a
reduction reaction when exposed to reducing sugars, changing color from green to brick-red or rusty-brown. This
reaction produces a copper thiocyanate precipitate, which can be used in titration. The solution turns orange or brick-red
as simple carbohydrates lose their reducing properties. However, the test does not detect complex carbohydrates like
starch unless they are heated or digested. In Barfoed’s test, glucose and fructose form a red precipitate within minutes,
confirming the presence of monosaccharides. In contrast, sucrose and lactose show no immediate reaction, indicating
they are disaccharides. This test distinguishes monosaccharides by reducing cupric acetate to cuprous oxide, forming a
brick-red precipitate. A reaction within two minutes confirms monosaccharides, while a slower reaction, taking about ten
minutes, suggests disaccharides.

aims to enhance understanding of carbohydrates by investigating their molecular structure, reactivity, and physical properties. It
seeks to:

1. Distinguish between monosaccharides and disaccharides based on their molecular structure.

2. Understand how the structure of carbohydrates influences their solubility properties.
3. Predict the reactivity of various carbohydrates when reacting with different oxidizing agents.
4. Learn the structural differences between aldose and ketose sugars.

LITERATURE CITED:

References
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Cardoso, V. C., Carvalho, L. V. C., Edvaldo Sabadini, & Sabadini, E. (2012). Carbohydrate research. Solubility of Carbohydrates in
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Sapkota, A. (2022b, September 17). Molisch Test- Definition, Principle, Procedure, result, uses. Microbe Notes.
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