Experiment # 3

Carbohydrates

Fehlings and Benedicts Test

Audrey Duran
Jackie Pinili
Estella Fermil
Dianne Belbider
Roxanne Birondo
Shaira Pacete
Airene Villanueva
BMLS-2G
Group 3
May 2, 2015

TABLE OF CONTENTS

Title Page

Table of Contents

ii

INTRODUCTION

Objectives

Result

Discussion/Interpretation

References

Introduction
Carbohydrates make up about half of biological substances and are one of the
main types of nutrients. They are the most important source of energy for our body.
Carbohydrates include table sugar, cellulose in plants and many more including some
components of important biomolecules like DNA. They are polyhydroxyaldehydes
(contains an aldehyde group) and polyhydroxyketones (contains a ketone group) and
may have carbon atoms ranging from as few as three to as many as thousands.
Monosaccharides are the simplest carbohydrates whose carbon atoms links only
range from about three to six. When these linkages combine through glycosidic bonds,
they form disaccharides, oligosaccharides and polysaccharides. The saccharide units
may assume linear or cyclic structures.
Many tests are used to detect specific carbohydrates, two of which are the Fehlings
and Benedicts tests. They are both tests that detect the presence of reducing sugars,
these reducing sugars include all monosaccharides and many disaccharides, including
lactose and maltose. This is because of the sugars ability to reduce the Copper (II) in
both the reagents to Copper (I), which shows up as a brick red precipitate signifying the
positive result. These two tests can also help one gauge the concentration of the
reducing sugar present, due to the fact that the intensity of the color is directly
proportional to the sugar concentration.

Fehlings Test and Benedicts Test

Objectives:
To detect the presence of reducing sugars through a visible positive result of
formation of brick red precipitate from each sample.
Samples
Glucose
Sucrose
Lactose
Fructose
Presentation of Results:

Observations
Red precipitate
Blue solution
Red precipitate
Red precipitate

*Color of reagent: Blue

Discussion/ Interpretations of Results:

BENEDICTS TEST and FEHLINGS TEST
a) What is the principle involved in Benedicts test and Fehlings test?
Fehlings test is a test for the detection of monosaccharides and and other reducing
sugars (Ex. Maltose). It gives a positive result for aldose monosaccharides because of
its oxidizable aldehyde group but also for ketose monosaccharides, because the
Fehlings reagent is able to convert ketoses to aldoses due to its base component.
Benedict's reagent is a chemical reagent commonly used to detect presence of reducing
sugar(s). This includes all monosaccharides and many disaccharides, including lactose
and maltose. Benedict's test will detect the presence of aldehydes, and alpha-hydroxyketones, including those that occur in certain ketoses. Thus, although the fructose is a
ketose and is not strictly a reducing sugar, it is an alpha-hydroxy-ketone, and gives a
positive test because it is converted to the aldoses glucose and mannose by the base in
the reagent.
b) What is the purpose of the tests?

The purpose of the test is to be able to detect reducing sugars and their
concentration if present.
c) What components make up the Fehlings reagent? The Benedicts reagent?
State the function of each component.
Fehling's solution contains copper(II) ions complexed with tartrate ions in sodium
hydroxide solution. Complexing the copper(II) ions with tartrate ions prevents
precipitation of copper(II) hydroxide.
Benedict's solution contains copper(II) ions complexed with citrate ions in sodium
carbonate solution. Again, complexing the copper(II) ions prevents the formation of a
precipitate - this time of copper(II) carbonate.
d) Provide the type of equation used in the tests

e) Show the sample equation involved in the reaction between the

Fehlings/Benedicts reagent and your sample/s which gave a positive result.

Samples

Observations

Glucose

Red-orange precipitate

Sucrose

Blue solution

Lactose

Red-orange precipitate

Fructose

Red-orange precipitate

Fehlings test:
Glucose and Fructose:

Lactose

Benedicts test:
Glucose: 2 CuSO4.5H2O + C6H12O6 -------> C6H12O7 + Cu2O + 2H2SO4 + 8 H2O
f) Provide a possible explanation for:

Positive result
Glucose, Fructose and Lactose solutions tested positive in both the Fehlings and the
Benedicts test as well as the urine (glucose containing) samples because they all
contain reducing sugars (lactose) and are aldoses (glucose) or monosaccharide
ketoses (fructose) which can be converted to aldoses by the reagent. Aldoses oxidize
due to their aldehyde functional groups and thus are able to reduce copper(II) to
copper(I), yielding a positive result (brick red precipitate).
Negative result
Sucrose was negative because both the anomeric carbons in its glucose and fructose
chains are tied up by its glycosidic bond, thus it is unable to oxidize and in turn reduce
the copper(II), yielding a negative result.
g) Why must the Fehlings reagent be freshly prepared?
It must be prepared fresh because it will undergo oxidation on standing (from the air)
and hence will not be effective.
h) Why is Fehlings test not used for the analysis of carbohydrates in clinical
samples?
Because Fehlings reagent is, unstable, is only a qualitative test, auto reduction of
cupric hydroxide occur resulting in false positive test.

i) State other advantages of Benedicts reagent over Fehlings reagent

1.

FEHLINGS TEST
The reagent is unstable

BENEDICTS TEST
1. The reagent is very stable

2. It has to be prepared in two parts

2.

Single solution is prepared and

which has to be stored separately

3. It is only a qualitative test

storage is
convenient.
3. It is both qualitative and semi-

4. The strong alkali (potassium

quantitative test
4. Since soda carbonate is a very

hydroxide) present in the reagent

weak base the destruction of

can destroy base the destruction of

carbohydrate is insignificant

carbohydrate
5. Auto reduction of cupric hydroxide

5.

Auto reduction doesnt take place

occur resulting in false positive test

6. It is more sensitive

6.

It is less sensitive

j) Differentiate between the 3 kinds of urine samples: fasting, post-prandial and

random. Which sample is the MOST suitable for use in a clinical laboratory test?
Explain your choice.
Fasting sample 12 hour urine without any food/drink ingestion.
Random just simple urine, without any did actions.
Post-prandial - The patient is given glucose water in a much diluted quantity and a
blood test is conducted after two to two and a half hours.
The fasting urine is the most suitable because it is the most natural urine that we
excrete and it is not yet mix with the chemicals etc. that we ingest.
k) What is Diabetes Mellitus? Differentiate between Type 1 and Type 2 diabetes. Is
it possible to have sugar diabetes even if no member of your family has it?
Explain your answer.
Diabetes mellitus is a kind of metabolic disease that is brought about by either the
insufficient production of insulin or the inability of the body to respond to the insulin
formed within the system.
What is diabetes mellitus type 1?

Diabetes mellitus type 1 is caused by the loss of beta cells found in the islets of
Langerhans in the pancreas.
Beta cells primarily produce insulin and hence, their loss will lead to huge insulin
deficiencies in the body.
In most cases, the loss of the beta cells is caused by an autoimmune attack mediated
by the bodys own T-cells.
Type 1 diabetes mellitus makes up 10% of all the diabetes mellitus cases in Europe and
in North America.
Up until today, there have been no preventive measures found for disease. Most of the
patients who acquire this type of diabetes are found in the younger populations.
Because of this, Type 1 diabetes mellitus is also now known as the juvenile diabetes.
What is diabetes mellitus type 2?
Another type of diabetes mellitus is the type 2 diabetes. Type 2 diabetes is generally
characterized by the bodys resistance to insulin. This is primarily attributed to the loss
of certain insulin receptors in the tissues that are supposed to mediate the entrance of
insulin into the bodys cells.
This particular kind of diabetes is the most common kind that afflicts most of the
reported cases of the disease. Type 2 diabetes usually leads to hyperglycemia which
can be treated by a series of medications that can either improve the bodys insulin
sensitivity or improve the livers production of insulin in the system.
The last major category of diabetes mellitus is the gestational diabetes. This type of
diabetes commonly occurs in pregnant women and may disappear or even progress
right after delivery. Gestational diabetes generally manifests the same kind of
complication found in type 2 diabetes.
Just like in type 2 diabetes, the body also exhibits a resistance to insulin and can
sometimes produce insufficient amounts of the substance as well. This kind of diabetes
is fully treatable but may require very close monitoring.

And it is still possible to have a sugar diabetes even is not inherited. DM is also
acquired when were engaging in a poor lifestyle and abandoned our self in eating too
much sweet.
l) Is one considered a diabetic if one obtains a positive result for sugar in a
fasting urine sample? Explain your answer.
No, because only the amylase activity is the one that is tested on the urine sample. The
test is not that accurate. Maybe the test only reacts with the reducing sugar and not with
the other sugar. Sugar can appear in the urine even if blood sugar isn't high. Also, sugar
in the urine doesn't reveal how giant blood sugar is. Sugar in the urine can tip a doctor
off that you may have diabetes, but it's a really outdated method of detecting and
monitoring diabetes.

References:
//www.scribd.com/doc/206925009/Carbohydrates
http://www.khalidshadid.com/uploads/3/9/2/0/3920808/carbohydrates_main.pdf
http://www.nku.edu/~whitsonma/Bio150LSite/Lab
%203%20Organic/Bio150LRevMolec.html