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Biochemistry lab3.pdf

The document outlines the Fehling test, a qualitative analysis method used to differentiate between reducing and non-reducing sugars in carbohydrates. It describes the preparation of Fehling's solution from two components, Fehling A and B, and details the procedure, expected results, and limitations of the test. The discussion includes insights on the chemical reactions involved and the reasons for keeping the solutions separate.

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1 views

Biochemistry lab3.pdf

The document outlines the Fehling test, a qualitative analysis method used to differentiate between reducing and non-reducing sugars in carbohydrates. It describes the preparation of Fehling's solution from two components, Fehling A and B, and details the procedure, expected results, and limitations of the test. The discussion includes insights on the chemical reactions involved and the reasons for keeping the solutions separate.

Uploaded by

hawraazaid04
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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PUNIVERSITY OF JABIR IBN HYYAN REPUBLIC OF IRAQ

COLLEGE OF PHARMACY MINISTRY OF HIGHER EDUCATION


3ED STAGE AND SCIENTIFIC RESEARCH

Biochemistry lab
Qualitative Analysis of Carbohydrates
Fehling Test

:BY
A5
‫حوراء زيد عدنان‬
‫زينب عماد خضير‬
‫فاطمة سلمان علي‬
‫ضحى عقيل عبد الكريم‬

SUPERVISOR :MSC.ALAA HUSSEIN


Introduction

Fehling’s solution, or Fehling’s reagent, is a chemical


reagent that is used to distinguish between
an aldehyde and a ketone, other than α-hydroxy
ketone. Practically, it is used for the determination of
reducing and non-reducing sugars that are present in
carbohydrates. The test employed for this
purpose is known as Fehling’s test.

Fehling's solution is still freshly formulated in the lab.


Initially, it was created as two separate solutions,
known as Fehling's A and Fehling's B. Fehling's A is
a blue aqueous solution of pentahydrate crystals of
copper(II) Sulphate, while Fehling's B is a transparent
solution of tartrate of aqueous potassium sodium
(also known as Rochelle salt) and a solid alkali
(commonly sodium hydroxide).

To get the final Fehling's solution, which is a deep


blue colour, equivalent amounts of the two mixtures
are blended together.
Tools

1/ 4 test tubes
2 for fehling’s reagent (A & b)
1 test tube for glucose and 1 for
fructose
2/ 4 pipttes
3/ Water path

Materials
1/ Glucose as a reducing suger
2/ Fructose as non-reducing suger
3/ Fehling reagent which consists of Fehling
A&B
Fehling A : Copper (ll) sulphate which is
blue colour

Fehling B: composed of Potassium sodium


tartrate (Rochelle salt) + strong alkali like
sodium hydroxide
which is a clear solution
Procedure

TAKE CONTROL OF 1 ML OF DISTILLED

WATER IN A DIFFERENT TEST TUBE

ADD 2-3 DROPS OF FEHLING’S REAGENT

INTO BOTH THE TEST TUBES AND MIX

THEM.

GIVE A WATER BATH TO BOTH THE TEST

TUBES FOR ABOUT 1-2 MINUTES.

OBSERVE THE COLOR CHANGE IN THE

TEST TUBES.
Results

POSITIVE RESULT: THE PRESENCE OF REDUCING SUGARS IS


INDICATED BY THE APPEARANCE OF REDDISH-BROWN
PRECIPITATE IN TEST TUBES.
NEGATIVE RESULT: THE LACK OF REDDISH PRECIPITATE OR THE
APPEARANCE OF A DEEP BLUE COLOUR INDICATES A NEGATIVE
RESULT AND A LACK OF REDUCING SUGARS.
Discussion

Q/ What happens when we add Fenling B to fehling A?


we will gain a complex due to reaction between CuSO4 and
potassium sodium tartrate following this equation bellow:

Q/ What is the limitation of this test?


1/ This test is unable to detect aromatic aldehydes due to the
resonance.
2/ The reaction requires an alkaline environment to take place.
If the environment is acidic, the copper ions will be stable and
do not get oxidized and fail the reaction.
Discussion

Q/ Why fehling A & B being separated?


Fehling's A and B solutions are kept separate
because the bistartratocuprate (II) complex
that is formed will easily degrade if they are
combined.

Q/ Why is Fehling solution blue?


The initial color of the solution is blue because it
contains copper ions.

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