Names: Bryle Kristiann Camarote Date Performed: February 19, 2014

Nimrod Romelo Date Submitted: February 26, 2014

Sarah Jane Valdon
Group #3

EXPERIMENT #8
Carbonyl Compounds and Carbohydrates

I. INTRODUCTION

A carbonyl group is a chemically organic functional group composed of a carbon atom

double-bonded to an oxygen atom. The simplest carbonyl groups are aldehydes and ketones
usually attached to another carbon compound. These structures can be found in many aromatic
compounds contributing to smell and taste.
A comparison of the properties and reactivity of aldehydes and ketones with those of
the alkenes is warranted, since both have a double bond functional group. Because of the
greater electronegativity of oxygen, the carbonyl group is polar, and aldehydes and ketones
have larger molecular dipole moments than do alkenes. The resonance structures on the right
illustrate this polarity, and the relative dipole moments of formaldehyde, other aldehydes and
ketones confirm the stabilizing influence that alkyl substituents have on carbocations (the
larger the dipole moment the greater the polar character of the carbonyl group). We expect,
therefore, that aldehydes and ketones will have higher boiling points than similar sized alkenes.
Furthermore, the presence of oxygen with its non-bonding electron pairs makes aldehydes and
ketones hydrogen-bond acceptors, and should increase their water solubility relative to
hydrocarbons.
The polarity of the carbonyl group also has a profound effect on its chemical reactivity,
compared with the non-polar double bonds of alkenes. Thus, reversible addition of water to the
carbonyl function is fast, whereas water addition to alkenes is immeasurably slow in the
absence of a strong acid catalyst. The carbonyl is polar due to the greater pi electron density at
the C=O bond. This property of the carbonyl group gives rise to a set of reactions characteristic
of aldehydes and ketones called the nucleophillic addition.
Carbohydrates are the most abundant class of organic compounds found in living
organisms. They originate as products of photosynthesis, an endothermic reductive
condensation of carbon dioxide requiring light energy and the pigment chlorophyll. They are
the major source of metabolic energy, both for plants and for animals that depend on plants for
food. Aside from the sugars and starches that meet this vital nutritional role, carbohydrates
also serve as a structural material (cellulose), a component of the energy transport
compound ATP, recognition sites on cell surfaces, and one of three essential components of
DNA and RNA.
 Carbohydrates consist of the elements carbon (C), hydrogen (H) and oxygen (O) with a ratio
of hydrogen twice that of carbon and oxygen. Carbohydrates include sugars, starches, cellulose
and many other compounds found in living organisms. In their basic form, carbohydrates are
simple sugars or monosaccharides. These simple sugars can combine with each other to form
more complex carbohydrates. The combination of two simple sugars is a disaccharide.
Carbohydrates consisting of two to ten simple sugars are called oligosaccharides, and those
with a larger number are called polysaccharides.
Their structure is composed of the functional groups, aldehyde and ketone, which are
attached with various amount of hydroxyl groups. The hydroxyl groups are usually attached to
the carbons not a part of the aldehyde or ketone functional groups, to form aldoses and
ketoses. Monosaccharides are the simplest carbohydrates in that they cannot be hydrolyzed to
smaller carbohydrates. They are aldehydes or ketones with two or more hydroxyl groups.
Monosaccharides are classified according to three different characteristics: the placement of
its carbonyl group, the number ofcarbon atoms it contains, and its chiral handedness. If the
carbonyl group is an aldehyde, the monosaccharide is an aldose; if the carbonyl group is
a ketone, the monosaccharide is a ketose.

Two joined monosaccharides are called a disaccharide and these are the simplest
polysaccharides. Examples include sucrose and lactose. They are composed of two
monosaccharide units bound together by a covalent bond known as a glycosidic linkage formed
via a dehydration reaction, resulting in the loss of a hydrogen atom from one monosaccharide
and a hydroxyl group from the other. The formula of unmodified disaccharides is C12H22O1.
Fructose is a 6-carbon polyhydroxyketone. It is an isomer of glucose; i.e., both have the
same molecular formula (C6H12O6) but they differ in structure. Crystalline fructose adopts a
cyclic six-membered structure owing to the stability of its hemiketal and internal hydrogen-
bonding.
Sucrose is the organic compound commonly known as table sugar and sometimes
called saccharose. A white, odorless, crystalline powder with a sweet taste, it is best known for
its role in food. The molecule is a disaccharide composed of
the monosaccharides glucose and fructose with themolecular formula C12H22O11.
In sucrose, the components glucose and fructose are linked via an ether bond between
C1 on the glucosyl subunit and C2 on the fructosyl unit. The bond is called a glycosidic linkage.
Glucose exists predominantly as two isomeric "pyranoses" (α and β), but only one of these
forms links to the fructose. Fructose itself exists as a mixture of "furanoses", each of which
having α and β isomers, but only one particular isomer links to the glucosyl unit. What is
notable about sucrose is that, unlike most disaccharides, the glycosidic bond is formed between
the reducing ends of both glucose and fructose, and not between the reducing end of one and
the nonreducing end of the other. This linkage inhibits further bonding to other saccharide
units. Since it contains no anomeric hydroxyl groups, it is classified as a non-reducing sugar.
Starch is the major form of stored carbohydrate in plants. Starch is composed of a
mixture of two substances: amylose, an essentially linear polysaccharide, andamylopectin, a
highly branched polysaccharide.
 Starches are transformed into many commercial products by hydrolysis using acids or
enzymes as catalysts. Hydrolysis is a chemical reaction in which water is used to break long
polysaccharide chains into smaller chains or into simple carbohydrates.

Cellulose is a polymer of β-D-Glucose, which in contrast to starch, is oriented with -

CH2OH groups alternating above and below the plane of the cellulose molecule thus producing
long, unbranched chains. The absence of side chains allows cellulose molecules to lie close
together and form rigid structures. Cellulose is the major structural material of plants. Wood is
largely cellulose, and cotton is almost pure cellulose. Cellulose can be hydrolyzed to its
constituent glucose units by microorganisms that inhabit the digestive tract of termites and
ruminants. Cellulose may be modified in the laboratory by treating it with nitric acid (HNO3) to
replace all the hydroxyl groups with nitrate groups (-ONO2) to produce cellulose nitrate
(nitrocellulose or guncotton) which is an explosive component of smokeless powder.

The compound 2,4-Dinitrophenylhydrazine can be used to qualitatively detect the

carbonyl functionality of a ketone or aldehyde functional group.
Tollens' reagent is a chemical reagent most commonly used to determine whether a
known carbonyl-containing compound is an aldehyde or a ketone. It is usually ammoniacal
silver nitrate, but can also be other mixtures, as long asaqueous diamminesilver(I) complex is
present. Once it has been ascertained that there is a carbonyl group on an organic
molecule using 2,4-dinitrophenylhydrazine (also known as Brady's reagent or 2,4-DNPH),
Tollens' reagent can be used to determine whether the compound is a ketone or an aldehyde.
Importantly, there is a special case in which Tollens' reagent will give a positive for a ketone; if
the ketone is an alpha-hydroxy ketone, then the Tollens' reagent will react.

The test rests on the premise that aldehydes are more readily oxidised compared with
ketones; this is due to the carbonyl-containing carbon in aldehydes having an attached
hydrogen. The diamminesilver(I) complex in the mixture is an oxidizing agent and is the
essential reactant in Tollen’s reagent.

In the Iodoform test, when iodine and NaOH are used as reagents, a positive reaction
gives Iodoform. Iodoform (CHI3) is a pale yellow substance. Due to its high molar mass due to
the three iodine atoms, it is solid at room temperature. It is insoluble in water and has an
antiseptic smell. A visible precipitate of this compound will form from a sample only when
either a methyl ketone, ethanol, a secondary alcohol or ethanol are present.
II. DATA AND DISCUSSION

In this experiment, various carbonyl compounds and carbohydrates were used to be tested
in different methods. These compounds are the aldehydes: benzaldehyde and acetaldehyde,
the ketones: acetone and cyclohexanone, the monosaccharides: glucose and fructose, and the
polysaccharides: starch and cellulose.

A. Solubility Behavior

COMPOUNDS OBSERVATIONS
Immiscible, clear liquid
Benzaldehyde Jelly-like benzaldehyde layer settled at the
bottom
Acetone Soluble, clear solution
Glucose Soluble, clear solution
Starch Soluble, clear solution

The first part of the experiment determines the solubility of the selected compounds with
water (H2O). As seen on the table above, benzaldehyde is immiscible with water. The main
reason for its immiscibility is its polarity. Benzaldehyde is a nonpolar monosubstituted benzene
ring. The ring has great pi electron density and since the carbonyl group substituent is an
electron withdrawing group, it affects the negativity of the ring making it less negative and
thereby reducing its polarity.
For acetone, its solubility with water showed a positive result (miscible). Acetone is a
polar compound due to its short chain R-group and its electron rich carbonyl group. Because
like dissolved like, this justifies its solubility. The acetone can form hydrogen bonds with water
in which the electron rich oxygen forms a dipole-dipole attraction with the partially positive
hydrogen of water.
Glucose and starch are also both soluble in water. Since they have many hydroxyl
groups (-OH), this makes them polar molecules and can form H-bonds with water and are
therefore soluble.
As we can see, their chemical structure contributed mainly to their solubility in a polar
substance (in our case, water). The ketone (acetone), comprises of a C=O group attached to an
alkyl group. The C=O group attracts electrons towards itself since oxygen is more
electronegative than the neighboring carbons while the alkyl group donates electrons to the
carbonyl group. This higher electron density near the oxygen atom results to a polar molecule.
On the other hand, the aldehyde (benzaldehyde), has a carbonyl group but is nonpolar.
Its carbonyl group is attached to the hydrophobic benzene ring, and this ring is already rich
in electron density. Since the carbonyl group attracts electrons, the electron density of the ring
decreases and its polarity is altered making it nonpolar. Glucose and starch are carbohydrates.
They both contain hydroxyl groups and these groups are electron withdrawing. This electron
withdrawing activity makes their molecules polar.
 In general, Benzaldehyde has an aromatic ring and a presence of a substituent. The ring has
a greater pi electron density while the substituent takes away electron from the ring making it
less negative. With this mechanism, it made the ring less polar and so benzaldehyde is insoluble
in water. Because of the greater C=O bond in the structure of the acetone, it is polar and is
soluble in water. The positive H atoms are attracted to one of the lone pairs of the oxygen in
acetone forming dipole-dipole attractions thus releasing energy which is sufficient in breaking
the water molecules and acetone bonds and so possible for dissolving with each other. Glucose
and starch are carbohydrates containing –OH which are hydrophilic making these
carbohydrates polar and so they form bonds with water making them soluble with it.
Moreover, Benzaldehyde contains hydrophobic aromatic ring which cannot form H-bonds
with water, acetone contains C=O making it polar. Since oxygen is more electronegative than
carbon, there is greater pi electron density at the oxygen end of the C=O making it polar and
carbohydrates and starch contains many –OH group which is polar and is soluble in water.

B. Hydrolysis

COMPOUNDS OBSERVATIONS
Sucrose Hydrolyzed
Starch Hydrolyzed
Cellulose Hydrolyzed

In hydrolysis, the compounds were mixed with water, HCl, heated in boiling water for 30
minutes and neutralized with NaOH. Hydrolysis involves breaking of polysaccharides down into
their constituent monosaccharide units. As we can see on the table above, all samples were
hydrolyzed.
Sucrose is a disaccharide and is broken down into one equivalent glucose and fructose.
Starch and cellulose are both polysaccharides that have hundreds of glucose units for starch
and thousands of D-glucose units for cellulose. Both are broken down into their components.
The characterization of sugars as reducing or non-reducing gives useful clues as to their
structures. Consider the disaccharides maltose and fructose. Maltose contains a hemiacetal
functional group and is a reducing sugar. In fructose, both anomeric carbons are in acetal
functional groups, so fructose is a non-reducing sugar.
 The linkages between the monosaccharide ring units in disaccharides are acetal
linkages. We can envision them as being made by the formation of an acetal from a hemiacetal
and an alcohol. For this purpose, the hemiacetal includes the anomeric carbon of a
monosaccharide and the alcohol role is played by a specific OH group of a second
monosaccharide. The formation of maltose from two molecules of glucose is an example of
this:

There are several intriguing features of this conversion. First, it is catalyzed by the
enzyme maltase. The term "catalyzed" implies that enzyme speeds up the reaction in both
directions, so that both formation and hydrolysis (conversion from acetal to hemiacetal using a
molecule of water) are faster with the enzyme. Enzymatic catalysis is usually also very specific.
In this case, that specificity shows up in the fact that the new acetal linkage has the alpha
configuration, not the beta (and correspondingly, maltase catalyzes the hydrolysis of an alpha
linkage but does nothing to the beta linkage). Also, only the OH group on the number four
carbon atom is used as the alcohol when others, such as the ones on carbons 1, 2, 3 and 6
might have been used. This suggests that the enzyme holds the two molecules of glucose in
specific positions so that only the OH on carbon 4 of one of the glucose can reach the anomeric
carbon of the other glucose.
Fructose provides an example of a disaccharide in which the acetal linkage joins the
anomeric carbons of a glucose molecule to the anomeric carbon of a fructose molecule. In this
case there is no hemiacetal functional group, so fructose is a non-reducing sugar.
Also, here one of the rings has five members rather than six, showing that the
cyclization of fructose from the open-chain form to the hemiacetal cyclic form uses the OH at
carbon 5 and the carbonyl carbon 2.
 We can also look more carefully at fructose. In its cyclic form the anomeric (hemiacetal)
carbon is involved in two carbon-carbon bonds. This means that when we open the molecule
up to its open chain form the anomeric carbon becomes a keto carbonyl group. Fructose is thus
an example of a ketose, a sugar in which the carbonyl group is a ketone rather than an
aldehyde.

If we now return to our first look at polysaccharides, we can see that amylose starch is
composed of many glucose monosaccharide units which are linked together by acetal
functional groups involving the anomeric carbon of one glucose and the number four carbon of
the next glucose. Repetition of this pattern many times gives the polymer.

Amylose is a linear polymer with few branches. In amylopectin, another type of starch,
there are branches which involve acetal linkages through the oxygen on carbon 6. Glycogen,
sometimes called animal starch, is a similar polymer found in animals as a storage medium for
glucose. Glycogen is even more highly branched than amylopectin.

Hydrolysis of starch involves the cleavage of the acetal functional groups with the
addition of a molecule of water for each acetal linkage and the production of many molecules
of glucose. This is done by the enzymes called glycosidases which are found in saliva. These
enzymes work only on alpha acetal linkages and do not attack beta linkages. Such beta linkages
are found in cellulose. Since our glycosidases are unable to hydrolyze the beta linkages in
cellulose, we cannot digest cellulose, even though it is also a polymer of glucose.
 Of course, there are enzymes which hydrolyze the beta linkages in cellulose. Such
enzymes are found in the bacteria which inhabit the stomachs of ruminants such as cattle and
sheep, which makes cellulose digestible by ruminants. They are also found in fungi which rot
wood.
The specificity of enzymes allows one monosaccharide, glucose, to be the building block
for both starch, which we think of as a major source of energy in our foods, and cellulose, which
we regard as a structural material in trees and a major component of paper. If we look at this in
the context of the use of these materials in a plant, starch is found as a storage medium for
glucose in seeds and tubers. It is used as a source of glucose both for energy and as a raw
material for cellulose as the plant sprouts and enters its initial growth period. Enzymes specific
for alpha linkages present in the sprouting plant hydrolyze the starch to glucose, as they do in
the malting process used in beer and whisky production.

The cellulose produced as the plant grows is a major structural component of the plant.
It must be quite stable if it is to serve that purpose, so enzymes specific for the alpha linkage do
not attack its beta acetal functional groups and it is not readily hydrolyzed. The small
stereochemical distinction between the alpha and beta linkages leads to very large
consequences in the chemistry and function of starch and cellulose.

C. Chemical Reactivity

1. Tollen’s Reagent
Compounds Observations
Acetaldehyde Soluble, clear
Silver mirror formed showing a positive test
(+)
Acetone Soluble, clear
No formation of Silver mirror showing a
negative test (-)
 2. Iodoform Test
Compounds Observations
Acetone Formation of yellow precipitate(+)
Cyclohexanone No formation of precipitate (-)
3. Reaction with 2,4-DNP
Compounds Observations
Acetone Orange precipitate(+)
Acetaldehyde Orange precipitate(+)

Tollen’s reagent was used to distinguish to determine the chemical reactivity of carbonyl
compounds such as acetaldehyde and ketones.
The difference between an aldehyde and ketone is the presence of a hydrogen atom
attached to the carbon-oxygen double bond in the aldehyde. Ketones, on the other hand, don’t
have that hydrogen. The presence of hydrogen atom makes aldehydes very easy to oxidize. Or,
put another way, they are strong reducing agents. Because ketones don’t have that particular
hydrogen atom, they are resistant to oxidation.
Meanwhile, Tollen’s reagent contains the diamminesilver(I) ion, *Ag(NH3)2++. This is
made from silver(I) nitrate solution. The addition of a drop of sodium hydroxide solution gives a
precipitate of silver(I) oxide, and then subsequently adding just enough dilute ammonia
solution redissolves the precipitate.

Aldehydes are very easily oxidized to yield carboxylic acids or their salts if the reaction is
done in basic media. Since ketones are not readily oxidized, this test is a useful method of
differentiating between aldehydes and ketones. The ammoniacal silver hydroxide solution used
in the Tollen’s Test is a very mild oxidizing agent (actually the Ag+1 ion). The silver ion is
reduced to metallic silver in a positive reaction. Sometimes a silver mirror forms on the test
tube. They are easily oxidizable because they react with Tollen’s reagent while ketones are not
oxidizable because it does not react with Tollen’s reagent.
 Ordinary ketones do not give a positive result in this test. The test should be used only if it
has already been shown that the unknown compound has a carbonyl group. It should be noted
that certain aromatic amines, phenols and amino ketones are also able to reduce ammoniacal
silver nitrate.
In the experiment, the addition of a few drops of the aldehyde or ketone was added to
the prepared reagent and was warmed gently in a hot water bath for minutes. It was observed
that in ketone, there was no change in the colorless solution; while in aldehyde, the colorless
solution produced a silver mirror on the walls of the test tube. This explains that aldehyde
reduces the diamminesilver(I) ion to metallic silver. Because the solution is alkaline, the
aldehyde itself is oxidized to salt of a corresponding carboxylic acid. The reaction of
acetaldehyde with Tollen’s reagent is:

The iodoform test is distinctive in the presence of methyl ketones. In the experiment,
only the acetone gave a positive result in which the appearance of a yellow precipitate
surfaced. Acetone has two methyl groups while cyclohexane does not. The removal of methyl
group of ketone from the molecule causes the production of iodoform (CHI3).

The compound 2,4-dinitrophenylhydrazine reacts with aldehydes and ketones to from

2,4-dinitrophenylhydrazones. Both acetone and acetaldehyde gave a positive result where the
formation of orange precipitate took place.

Methyl ketones can be distinguished from other ketones by their reaction with iodine in
a basic solution to yield iodoform (CHI3) as a yellow colored precipitate. However, acetaldehyde
(CH3CHO) and alcohols with their hydroxyl groups at the 2 position of the chain will also form
iodoform under these same conditions. Alcohols of the type described are easily oxidized to
methyl ketones under the conditions of the iodoform reaction (I2 is an oxidizing agent). The
other product of the iodoform reaction is the sodium or potassium salt of a carboxylic acid.

Both ethanol and 2-propanol are oxidized by iodine to give ethanal or acetone (1).
 When a - methyl carbonyl compounds react with iodine in the presence of a base, the
hydrogen atoms on the carbon adjacent to the carbonyl group (a hydrogens) are subsituted by
iodine to form tri iodo methyl carbonyl compounds which react with OH - to produce iodoform
and carboxylic acid (2):

*Reaction mechanism:
The hydrogen atoms on the methyl group are slightly acidic and can be removed with
hydroxide. The carbanion formed then reacted with iodine molecules to give a iodide ion and a
monoiodonated methyl carbonyl derivate. Introduction of the first iodine atom (owing to its
electronegativity) makes the remaining hydrogens of the methyl group more acidic. Hence a
base-catalized iodination of a monohalogenated methyl carbonyl derivate occurs at the carbon
that is already substituted. Finally a triiodomethyl carbonyl derivate is formed.

The next step is a nucleophilic attack by hydroxide on the carbonyl carbon atom. A
carbon - carbon bond cleavage occurs and a triiodomethanide ion departs. The
triiodomethanide ion is unusually stable. Its negative charge is dispersed by the three negative
iodine atoms. In the last step a proton transfer takes place between carboxylic acid and
triiodomethanide ion to form ultimately carboxylate ion and iodoform.
This implies that there is a presence of the carbon-oxygen double bond in the reaction. The
reactions involved are:

2- propanol + 2,4-DNP No reaction

Acetone + 2,4-DNP 2, 4 dinitrophenylhydrazone +H20
1-pentanol + 2,4-DNP No reaction
Glucose + 2,4-DNP 2,4 dinitrophenylhydrazone + H20

In general, in order for the carbonyl compounds to undergo haloform reaction, they
needed to have at least one methyl group. For example, in iodoform test, ketone with at least
one methyl group is oxidized and converted to yellow precipitate CHI3.

One methyl group is required for the carbonyl compounds to undergo haloform
reaction. Carbonyl compunds possessing the structure can also undergo the haloform reaction.
The bond between the carbonyl and methyl is cleaved to give a carboxylate ion and a haloform:

O O
+ 3X2 + 4OH- + CHX3 + 3X- + H2O
-
R CH3 R O
 D. Color Reactions of Carbohydrates

Carbohydrates are widely prevalent in the plant kingdom, comprising the mono-, di-,
oligo-, and polysaccharides. The common monosaccharides are glucose, fructose, galactose,
ribose etc. The disaccharides, i.e., the combination of two monosaccharides include sucrose,
lactose and maltose. Starch and cellulose are polysaccharides consisting of many
monosaccharide residues. Cellulose is the most abundant organic compound on this planet
since it forms part of the cell wall in plants.

Aldehydes (–CHO) and ketones ( = CO) are active groups in carbohydrates.

Carbohydrates contain many hydroxyl groups as well. The number of hydroxyl groups varies
with the number of carbon atoms. Monosaccharides contain the free aldehyde or ketone
group. Some disaccharides have the free aldehyde group (maltose) and some do not have the
free ones (sucrose). The polysaccharides, starch and cellulose, are polymers of
monosaccharides linked through the active groups.

The chemical properties of saccharides vary depending upon the number of hydroxyl
groups and the presence or absence of –CHO/= CO groups. These variations are the basis in the
development of color reactions to identify the saccharides.

Molisch Test
Compounds Observations
Cyclohexanone (-) brownish orange
Glucose (+) very light brown
Starch (+) very light brown
Sucrose (-) purple
Benedict’s Test
Compounds Observations
Glucose Red-orange(+)
Fructose Red-orange(+)
Sucrose Light blue solution(-)
Lactose Red-orange(+)
Sucrose hydrolysate Brick red(+)
Starch hydrolysate Brick red(+)
No precipitate, purple colored
Cellulose hydrolysate
solution(-)
 Osazone Formation
Compounds Time of Appearance
Glucose 05:20 min or 320 seconds
Fructose 01:45 min or 105 seconds
Sucrose
Lactose 01:15 min or 75 seconds

The Molisch test is a general test for the presence of carbohydrates. Molisch reagent is a
solution of alpha-naphthol in 95% ethanol.

Molisch's Test (named after Austrian botanist Hans Molisch) is a sensitive chemical test
for the presence of carbohydrates, based on the dehydration of the carbohydrate by sulfuric
acid to produce an aldehyde, which condenses with two molecules of phenol (usually α-
naphthol, though other phenols (e.g. resorcinol, thymol) also give colored products) resulting in
a red- or purple-colored compound.
The test solution is combined with a small amount of Molisch's reagent (α-naphthol
dissolved in ethanol) in a test tube. After mixing, a small amount of concentrated sulfuric acid is
slowly added down the sides of the sloping test-tube, without mixing, to form a bottom layer. A
positive reaction is indicated by appearance of a purple ring at the interface between the acid
and test layers.
In the presence of concentrated sulfuric acid, glycosidic linkages are hydrolyzed to give
monosaccharides.

nH2O + (C6H10O5)n  nC6H12O6

The rate of hydrolysis depends on the solubility of the carbohydrate in water. The
monosaccharides formed are then dehydrated to furfural (or hydroxymethylfurfural) and other
colored decomposition products.

Furthermore, Molisch test is useful for identifying any compound which can be
dehydrated to furfural or hydroxymethylfurfural in the presence of H2SO4. Furfural is derived
from the dehydration of pentoses and pentosans, while hydroxymethylfurfural is produced
from hexoses and hexosans.

Oligosaccharides and polysaccharides are hydrolyzed to yield their repeating monomers

by the acid. The alpha-naphthol reacts with the cyclic aldehydes to form purple colored
condensation products. Although this test will detect compounds other than carbohydrates (i.e.
glycoproteins), a negative result indicates the ABSENCE of carbohydrates.
 Since glucose and starch are carbohydrates, both of them gave a positive result. On the
other hand, cyclohexanone gave a negative result since it is a carbonyl compound and not a
carbohydrate. But sucrose gave a negative result. Sucrose is a carbohydrate and must therefore
be positive, but this error may be due to some factors such as contamination.

On the other hand, Benedict's test distinguishes reducing sugars from non-reducing
sugars.Glucose, fructose, and lactose have hemiacetal groups, so they are reducing sugars.
Hemiacetal groups are compounds that are derived from aldehydes and ketones respectively.
The Greek word hèmi means half. These compounds are formed by formal addition of
an alcohol to the carbonyl group.They gave a positive test, that is the red-orange color.

Sucrose was identified as nonreducing and it supported when it gave a negative test. In
the test, Cu2+ is reduced to Cu+ and the reducing sugar is oxidized to a carboxylic acid. The
general reaction is shown below:

Hydrolysates are products of hydrolysis which means that they are already converted to
monosacchardies. The hydrolysates of sucrose are glucose and fructose so they gave a positive
test for reducing sugars. On the other hand, hydrolysates of starch and cellulose didn’t give a
positive test. This means that they are nonreducing sugars.
The reaction between hydrolysates of starch and the hydrolysates of sucrose with
Benedict’s reagent differs in color. The later has a greenish tint than the former. Both have
reddish precipitates. Reducing sugars are usually detected by Benedict’s reagent, which
contains copper (II) ions in alkaline solution with sodium citrate added to keep the cupric ions in
the solution. The alkaline conditions of this test causes isomeric transformation ketoses to
aldoses. It results in all monosaccharides and most disaccharides reducing the blue cupric ion to
cuprous oxide (Cu2O) which is a brick red precipitate.

*Osazone formation differentiates reducing sugars from nonreducing sugars. Sugars

containing aldehydes, ketones, or hemiacetal groups are able to reduce an oxidizing agent and
therefore classified as reducing sugars. Without one of these groups, it is a nonreducing sugar.

Reducing sugars may also be detected and differentiated from each other by their
reaction with phenylhydrazine. The reaction is more extensive than the formation of
phenylhydrazones from simple carbonyl compounds. The products of the reaction are called
osazones. These are yellow, crystalline solids with well – defined melting points and crystalliine
structure. They are useful in the identification of simple sugars.

O 2N O 2N

O H2N N + H2O
+ NH NO 2 NH NO 2

Aldehyde 2,4-DNP 2,4-dinitrophenylhydrazone

or ketone
*Osazones are a class of carbohydrate derivatives formed by the reaction of a sugar
with phenylhydrazine. An osazone is a solid derivative of a sugar containing two
phenylhydrazone moieties. In theory, it should be possible to use the melting point of this
derivative to identify the unknown sugar.
In practice, this is not easily accomplished because osazone derivatives melt over a very wide
range and the identical osazone is obtained for more than one sugar. D-glucose, D-mannose,
and D-fructose all give the same osazone, so the melting point of the osazone could not
distinguish between sugars.
However, careful observation of the rate at which the osazone forms and the
appearance of the precipitate can differentiate between epimeric sugars. Osazones form at
different rates for different sugars: fructose reacts very rapidly, while glucose takes longer to
react. The appearance of the precipitate can also be different. The crystal structure ranges from
coarse (for glucose) to very fine (for arabinose).

1
.
 Monosaccharides such as glucose and fructose and disaccharide such as lactose are
reducing sugars. This is evident on the formation of yellow crystalline osazones upon reaction
with phenylhydrazine.

Sucrose, a nonreducing double sugar of glucose and fructose, gives no osazone. Because
sucrose doesn’t have a hemiacetal group, it is not in equilibrium with the readily oxidized
aldehyde or ketone. And thus, no osazone is formed.

Generally, Carbohydrates with the presence of even small amount of the open – chain form
allows the reactions associated with the carbonyl group to take place. Moreover, in the open –
chain form, the interaction between the carbonyl group and the ajacent hydroxyl group makes
the carbonyl group more susceptible to oxidation and more reactive to nucleophilic reagents.

Carbohydrates that can be hydrolyzed to two monosaccharide units are called

disaccharides. The monosaccharide units in disaccharides are joined by a glycosidic linkage
(specifically, an acetal or ketal linkage), which is really an ether linkage.

*Simple chemical tests are shown below to differentiate between the following compounds.
Equations for every reaction are also showed.

a. butanone and butanal (butyraldehyde)  Tollen’s Reagent

Butanone + Ag(NH3)2 no reaction
Butanol + Ag(NH3)2 Ag+ (s)

b. 2-propanol and acetone  2,4 -DNP

2 propanol + 2,4-DNP No reaction
Acetone + 2,4-DNP 2, 4 dinitrophenylhydrazone + H2O

c. Glucose and Butanal  Molisch Reagent

Butanol + Molisch reagent no reaction
Glucose + Molisch reagent nC6H1206

d. Sucrose and lactose  Benedict’s Reagent

Sucrose + Benedict’s reagent No reaction
Lactose + Benedict’s reagent +Cu20 (red ppt)

e. glucose and 1-pentanol  2,4- DNP

1 pentanol + 2,4-DNP No reaction
Glucose + 2,4-DNP 2,4 dinitrophenylhydrazone + H20
III. CONCLUSIONS

Aside from carbon and hydrogen, oxygen is also frequently found in important organic
compounds. Important groups of oxygen – containing compounds are the carbonyl
compounds. Those are compounds that contain the carbonyl group, C = O. They can be
classified as aldehyde or ketones depending on what groups are bonded to the C = O group.
If the compound has an aromatic ring and a presence of a substituent, the ring has a
greater pi electron density while the substituent takes away electron from the ring making it
less negative. With this mechanism, it made the ring less polar became insoluble in water.
Carbohydrates contain –OH which are hydrophilic that makes these carbohydrates
polar and form bonds with water making them soluble with it. The characterization of sugars as
reducing or non-reducing gives useful clues as to their structures.
Hydrolysis of starch involves the cleavage of the acetal functional groups with the
addition of a molecule of water for each acetal linkage and the production of many molecules
of glucose. Stereochemical distinction between the alpha and beta linkages leads to very large
consequences in the chemistry and function of starch and cellulose.
2 ,4-Dinitrophenylhydrazone Test are useful in distinguishing an aldehyde or ketone
from other functional groups. The solid products resulting from these reagents are also used as
solid derivatives for the identification of specific aldehydes and ketones.
Tollen’s Test is a useful method of differentiating between aldehydes and ketones since
aldehydes are easily oxidized while ketones are not.
Iodoform Test is used to distinguish Methyl ketones from other ketones by their
reaction with iodine in a basic solution to yield iodoform (CHI3) as a yellow colored precipitate.
Molisch's Test (named after Austrian botanist Hans Molisch) is a sensitive chemical test
for the presence of carbohydrates, based on the dehydration of the carbohydrate by sulfuric
acid to produce an aldehyde, which condenses with two molecules of phenol resulting in a red-
or purple-colored compound.
The Benedict's test allows us to detect the presence of reducing sugars (sugars with a
free aldehyde or ketone group). A positive test is indicated by the reduction of the Cu2+ ion to
Cu (I), (Cu2O) which is red brick in color and insoluble.
An osazone is a solid derivative of a sugar containing two phenylhydrazone moieties. In
theory, it should be possible to use the melting point of this derivative to identify the unknown
sugar. In practice, this is not easily accomplished because osazone derivatives melt over a very
wide range and the identical osazone is obtained for more than one sugar.

IV. REFERENCES

Peter Keusch. 2003. Iodoform Reaction: Nucleophilic Carbonyl alpha-Substitution, Test for the alpha-
Methyl Carbonyl Group.
Bill Kelly. 1998. ORGANIC II LABORATORY: Aldehydes and Ketones.
Bruce Knauer. 2004. Experiment #9: Identification of Aldehydes and Ketones.
McMurry, Organic Chemistry, 7th Philippine Edition, 2004.