Carbohydrates

post laboratory conference

Discussion Outline
1 3
Monosaccharides
- Fischer’s Formula &
Types of Sugars
- Reducing & Non-reducing
Haworth’s Formula sugars
- Glucose, Galactose, &

4 Tests for
Fructose

Carbohydrates
2 Disaccharides
- Maltose, Lactose, &
Sucrose
- Molisch’s, Fehling’s,
Benedict’s, & Barfoed’s Tests
Carbohydrates
» A biomolecule consisting of carbon,
hydrogen and oxygen atoms

Monosaccharides
» Glucose, Galactose, & Fructose

Structural Formula
» Fischer’s Formula & Haworth’s Formula
Carbohydrates
» A biomolecule consisting of carbon,
hydrogen and oxygen atoms

Monosaccharides
» Glucose, Galactose, & Fructose

Structural Formula
» Fischer’s Formula & Haworth’s Formula
Monosaccharides
Aldohexose
» Glucose and Galactose
» Aldehyde group (C1)
» Reducing sugars

Ketohexose
» Fructose
» Ketone group (C2)
» Reducing sugars
Disaccharides
Maltose
» Glucose + Glucose
» Reducing sugar

Lactose
» Glucose + Galactose
» Reducing sugar

Sucrose
» Glucose + Fructose
» Non-Reducing sugar
Types of Sugars
Reducing Sugars
» All monosaccharides, Maltose &
Lactose
» Carbohydrates that are reducing
sugars have a free carbonyl
(aldehyde or ketone) group

Non-Reducing Sugars
» Sucrose
» Carbohydrates that are non-reducing
sugars do not have a free carbonyl
(aldehyde or ketone) group
Molisch’s Test
Molisch’s test
 » To detect the presence of
carbohydrates

Purpose » Positive on all carbohydrates

Purpose
except trioses and tetroses

» Monosaccharides react faster than

disaccharides and polysaccharides
Glycoproteins Nucleic acids
 1.) Molisch’s Reagent

» A solution of ∝-naphthol in

Reagents
ethanol
» responsible for condensing with

Reagents
furfural derivative

2.) Sulfuric Acid (H2SO4)

» Responsible for dehydrating the

carbohydrate
 1.) Place 2 ml of sample in test
tube

2.) Place 2 ml of water in

another test tube

3.)

Procedure
Add 2-3 drops of Molisch’s
Reagent to both solutions

Procedure 4.)

5.)
Mix both solutions well

Pipette 1 ml conc. Sulfuric acid

along the side of both tubes

6.) Observe purple ring if sample

contains a carbohydrate
 » Carbohydrate gets dehydrated
by sulfuric acid

Principle
Principle » Furfural derivatives condense
with α-naphthol resulting in a
purple complex
Equation
Equation
 » Given the samples: glucose,

Application liver extract, and

Application
carrageenan, which of these
will yield a positive visible
result?
» Glucose gives a positive
result

» Liver extract yields a

positive result due to
glycogen

» Carrageenan yields a
positive result due to
galactose units
» Glucose gives a positive
result

» Liver extract yields a

positive result due to
glycogen

» Carrageenan yields a
positive result due to
galactose units
» Glucose gives a positive
result

» Liver extract yields a

positive result due to
glycogen

» Carrageenan yields a
positive result due to
galactose units
» Glucose gives a positive
result

» Liver extract yields a

positive result due to
glycogen

» Carrageenan yields a
positive result due to
galactose units
Fehling’s Test
Fehling’s test
 » The aldehyde group of sugar is
oxidized by copper

Principle ions to form acid.

Principle » Presence of reducing sugar =

formation of red precipitate of
cuprous oxide
Purpose » Detection of reducing sugars in a

Purpose
given solution
 1.) Copper (II) sulfate pentahydrate
(CuSO4.5H2O)

» responsible for oxidizing the

reducing sugar. 

Reagents 2.) Sodium Potassium

Reagents Tartrate/Rochelle salt

(NaK(C4H4O6) mixed with
Sodium hydroxide

» alkaline medium

» serves as a chelating agent in

the solution
 1.) Add sample in test tube

2.) Place water in another test

tube

Procedure 3.) Fehling’s solution A and B to be

Procedure
added in all the tubes

4.) Keep tubes in boiling water bath

5.) Observe and record if any

development of red precipitate
takes place
 » Given the following

Application samples, glucose, sucrose,

Application
fructose, and lactose, only
sucrose yields to a negative
result.
Equation » RCHO + 2 Cu2+ + 5 OH− →

Equation RCOO− + Cu2O + 3 H2O

 » This test is not used in analyzing
carbohydrates for it deteriorates
quickly

» It has caustic/corrosive properties

Application
Application » It is made up of two separate
solutions to be mixed together

» The Benedict’s Test is considered to

be more stable.
Benedict’s Test
Benedict’s test
 » Heating of reducing sugar

Principle solution with

Principle
Benedict’s reagent forms
visible positive results
 » Determine presence or
absence

Purpose
of reducing sugar(s) in a
solution

Purpose » Quantitatively determine

solution’s glucose
 1.) CuSO4.5H2O (Copper sulfate
pentahydrate)
» Provides cupric ions

2.) Na2CO3 (Sodium carbonate)

Reagents » Provides alkaline environment

Reagents 3.) Na3C6H5O7 (Sodium citrate, in

distilled water)

» Prevents precipitation of
cupric ions

» Can be kept longer during

storage
 1.) Add 1 ml of sample into a
clean test tube

2.) Add 2 ml of Benedict’s reagent

into same test tube

Procedure 3.) Mix the solution

Procedure 4.) Clamp your test tube and place

in a boiling water bath for 3-5
minutes

5.) Observe for color change/precipitate

formation
 » Given the following samples:
glucose, sucrose, fructose, lactose,
fasting urine, random urine, and
diabetic urine

»
Application
Glucose, Fructose, Lactose
Fasting urine, Diabetic urine will

Application
yield a positive result

» Sucrose yields a negative result

» Random urine sample may yield a

negative result
Equation
Equation
Equation
Equation
 » Clear indication of glucose
concentration

Advantage based on color of visible result

Advantage » Accessible and easy, long

storage
 1.) Fasting
» the first morning specimen or 8-
hour specimen
» has relatively higher levels of
cellular elements

Urine
2.) Post-prandial
» collected 2 hours after eating a

Samples
meal

» detect early symptoms of

prediabetes and diabetes types I
and II

3.) Random
» can be collected any time

» readily sent to a laboratory for

analysis since it’s easy to obtain
 »
Urine
Fasting urine sample is
best suited for use in a

Samples clinical laboratory test

 » Is one considered a diabetic if
one obtains a positive result for
sugar in a random urine

Application sample?

Application » A positive result of a random

urine sample does not
automatically indicate diabetes
Barfoed’s Test
Barfoed’s test
 »
Principle
Cupric ions are reduced to
cuprous ions

Principle » Reducing sugars are oxidized

Purpose » Distinguish monosaccharides

Purpose from disaccharides

 1.) Acetic Acid
» Serves as the acidic medium

Reagents
Reagents 2.) Copper Acetate
» Provides the copper (II)
ions
 1.) Add 1 ml of sample into a
clean test tube

2.) Add 1 ml of distilled water in

another

Procedure
3.) Add 2 ml of Barfoed’s Reagent
in each tube

Procedure 4.)

5.)
Mix the solutions

Heat the solutions in a boiling water

bath for 2-3 mins.

6.) Observe for the color and development

of brick-red precipitate in the solution
 » Given the following samples:
glucose, sucrose, fructose, and

Application
lactose, which of these will
yield a positive visible result?

Application » Glucose and Fructose will yield

a positive result
Equation
Equation
Visible
Results Positive Visible Result = brick-red precipitate
Negative Result = blue solution
 » Why is the heating of the
samples limited to a few

Application
minutes only?

Application » Prolonged heating on a

negative sample will yield a
false positive
 » Karki, G. (2018). Barfoed’s Test: Objective,
Principle, Reagents, Procedure and Result.
Retrieved from:
https://www.onlinebiologynotes.com/barfoeds-tes
t-objective-principle-reagents-procedure-and-resu
lt/

References » Barfoed’s Test. (n.d.). Retrieved from:

References
https://www.chemistrylearner.com/barfoeds-
test.html

» (n.d.). Barfoed’s Test. Retrieved from:

http://dept.harpercollege.edu/chemistry/chm/100/
dgodambe/thedisk/carbo/barf/barfoed.htm#:~:text
=Reactions%3A,so%20at%20a%20slower%20rat
e
.
 » Molisch’s Test. (n.d.). Retrieved November 10,
2020., from Byju’s The Learning App:
https://byjus.com/chemistry/molischs-test/

» Fehling's Solution - Formula, Preparation, Uses,

References
Fehling's Test (2020). Retrieved from
https://byjus.com/jee/fehlings-solution/#fehlings-t

References
est-procedure

» Fehling’s test (Principle, reagents and materials

required, procedure and result interpretation) -
Online Science Notes. (2019). Retrieved from
https://onlinesciencenotes.com/fehlings-test-princ
iple-reagents-and-materials-required-procedure-a
nd-result-interpretation
/
 » Fehling’s Solution: Definition, Example, and
Mechanism. (2020). Retrieved from
https://www.chemistrylearner.com/fehlings-soluti
on.
html

» solution, F., & Monica, P. (2016). Functional

References
difference of Benedict's solution and Fehling's
solution. Retrieved from

References
https://chemistry.stackexchange.com/questions/62
034/
functional-difference-of-benedicts-solution-and-fe
hlings-solution