Kate Coleen D.

Galera BS in Chemistry II April 24,2015

Experiment 10
Carbonyl Compounds and Carbohydrates

I.Data and Results

A. Solubility Behavior
Compound Homogeneity Observations
Benzaldehyde Heterogeneous Immiscible, clear liquid Jelly-like
benzaldehyde layer settled at the bottom
Acetone Homogenous Soluble, clear solution
Glucose (solid) Homogenous Soluble, clear solution
Starch (solid) Homogenous Soluble, clear solution
Cellulose (solid) Homogenous Insoluble

B. Hydrolysis of Di- and Polysaccharides

Compound Homogeneity Observations
Sucrose Homogenous Formed a neon pink solution with Hydrolyzed
small settled precipitates
Starch Homogenous Formed light pink solution Hydrolyzed
Cellulose Heterogeneous Formed gelatinous pink solution Hydrolyzed

C. Chemical Reactivity of Carbonyl Compounds

C.1. Reaction with Tollen’s reagent
Compound Observations
Acetaldehyde Formed a colorless solution with small black particles to gray
solution with small black particles. Soluble
Acetone Formed a colorless solution with small black particles but less than
acetaldehyde. Soluble

C.2 Iodoform Test

Compound Drops to undergo color change Observations
Acetone 2 Formed clear solution
Cyclohexane 3 Formed clear solution at the bottom
and oily yellow emulsion at the
upper layer.
Acetaldehyde 4 Formed clear solution

C.3 Reaction with 2, 4 – DNP

Compound Observations
Acetone Formed yellow solution with yellow orange
precipitates
Acetaldeyde Formed yellow-orange solution with orange
precipitates
 Glucose Formed only orange solution with no precipitates
Fructose Formed only orange solution with no precipitates
Cyclohexanone Formed yellow solution with neon yellow
precipitates
Benzaldehyde Formed yellow orange solution with yellow
orange precipiates

D. Color Reactions of Carbohydrates

D.1 Molisch Test
Compound Homogeneity/ Adding Molisch Adding concentrated H2SO4 Result
Solubilty Reagent
Starch Homogenous/ Formed black small Formed 3 layers, colorless at
Soluble precipitates the top, violet ring at the
center and pale red-violet at
the bottom. +
Heterogeneous mixture with
small black precipitates
Sucrose Homogenous/ Formed black small Formed dark brown yellow
Soluble precipitates solution with black
precipitates. Homogenous +
Glucose Homogenous/ Formed black small Formed brown solution with +
Soluble precipitates precipitates. Homogenous
Cyclohexanone Insoluble Formed oily solution Formed 2 layers, light brown
with orange color at at the top and brownish re at
the top and clear the bottom. Heterogeneous. _
solution with small
black precipitates at
the bottom.

D.2 Benedict’s Test

Compound Observations before
heating(Benedict’s reagent)
Sucrose (Hydrolates) Cloudy pinkish solution
Starch (Hydrolates) Two layers, purple at top and
indigo at the bottom
Cellulose (Hydrolates) Gelatinous violet solution
Lactose Sky blue solution
Glucose Sky blue solution
Fructose Sky blue solution
Observations after heating
Brown solution with no
precipitates. Soluble
Cloudy orange solution with no
precipitates. Soluble
Gelatinous pink solution with
dark pink around it and white
gelatinous small area under.
Sticky Texture. Soluble
Cloudy dark orange solution
with no precipitates. Soluble
Cloudy dark orange solution
with no precipitates. Soluble
Cloudy dark brown solution
with no precipitates. Soluble
 Sucrose Sky blue solution Cloudy light brown solution
with no precipitates. Soluble

D.3 Osazone Formation

Compound Time that precipitates formed Observations
Fructose 1 minutes and 12 seconds Formed pale yellow solution
with insoluble white particles
settled in.
Lactose After 5 minutes no precipitate Formed pale yellow ssolution
formed with insoluble white particles
settled in.
Sucrose After 5 minutes no precipitate Formed pale yellow solution
formed with insoluble white particles
settled in.
Glucose 3 minutes and 57 seconds Formed pale yellow solution
with insoluble white particles
settled in.

II.Discussions
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.
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.

A. Solubility Behavior
The first part of the experiment determines the solubility of the selected compounds with water.
Benzaldehyde is immiscible with water. The main reason for its immiscibility is its polarity. Benzaldehyde
is a nonpolar mono-substituted 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. Cellulose is
a huge polysaccharide polymer, it hydrogen bonds pretty much with water but the molecule is just too
big to be completely soluble. This bonding makes it insoluble in either polar or non-polar solvents.
In general, 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 of Di- and Polysaccharides

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. Starch, sucrose and cellulose were all 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.
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 glycosidases are
unable to hydrolyze the beta linkages in cellulose, it cannot digest cellulose, even though it is also a
polymer of glucose.
C.1. Reaction with Tollen’s reagent
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.
Theoretically, a ketone, will not undergo change in its colorless solution; while in aldehyde, the
colorless solution will produce a silver mirror on the walls of the test tube. This explains that aldehyde
reduces the diamine silver(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:

C.2 Iodoform Test

The iodoform test is distinctive in the presence of methyl ketones. Theoretically, Acetone will give
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 experimenter however obtained colorless solution upon dropping
10% NaOH.

C.3 Reaction with 2, 4 – DNP

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
yellow-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 o xidized to methyl ketones under the
conditions of the iodoform reaction where I2 is an oxidizing agent. The other product of the iodoform
reaction is the sodium or potassium salt of a carboxylic acid.

D. Color Reactions of Carbohydrates

D.1 Molisch Test
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 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. 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.

n H2O + (C6H10O5) n n C6H12O6

The rate of hydrolysis depends on the solubility of the carbohydrate in water. The monosaccharides
formed are then dehydrated to furfural 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, 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.

2. Benedict’s Test
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. 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.
Theoretically, sucrose is identified as non-reducing 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
monosaccharides. The hydrolysates of sucrose are glucose and fructose so they will give a positive test for
reducing sugars. On the other hand, hydrolysates of starch and cellulose will not give a positive test. This
means that they are non-reducing 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.

C.3 Osazone Formation

Osazone formation differentiates reducing sugars from non-reducing 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.

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).

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 non-reducing 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.

III.Conclusion
Carbonyl Compounds an important group of oxygen – containing compounds are the carbonyl
compounds – those that contain the carbonyl group, C = O. They are classified as aldehyde or ketones
depending on what groups are bonded to the C = O group.Carbohydrates are polyhydroxy aldehydes or
ketones and the derivatives or compounds which yield such on hydrolysis. Many of them are represented
by the general formula Cx(H2O)y, which led early workers to regard them as hydrates of carbon, hence
the term "carbohydrates”
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.
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
McMurry, John. Organic Chemistry. Thomson Publications. 2004
Peter Keusch. 2003. Iodoform Reaction: Nucleophilic Carbonyl alpha-Substitution, Test for the
alphaMethyl Carbonyl Group.
Bill Kelly. 1998. ORGANIC II LABORATORY: Aldehydes and Ketones.
Bruce Knauer. 2004. Experiment #9: Identification of Aldehydes and Ketones

V. Answers to Questions
1. Explain the solubility behavior of the samples used based on their chemical structures.
- 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.
- . Cellulose is a huge polysaccharide polymer, it hydrogen bonds pretty much with water but the molecule
is just too big to be completely soluble. This bonding makes it insoluble in either polar or non-polar
solvents.

2. What is the structural requirement for the haloform reaction of the carbonyl compounds?
- 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.

3. How would you rate the oxidizability of aldehydes and ketones: easily oxidizable, oxidizable, not
oxidizable? Justify your answer using specific chemical tests.
Aldehydes and ketones both have, as their functional group, a carbon-oxygen double bond (carbonyl
group). Accordingly, they undergo similar reactions. With the same reagent, aldehydes usually react faster
than ketones, mainly because there is less crowding at the carbonyl carbon. Aldehydes are also more
easily oxidized than ketones. Aldehydes are easily oxidized to acids which have the same number of
carbon atoms. An analogous reaction is not possible for ketones, whose oxidation to an acid requires the
rupture of a carbon-carbon bond and results in two acids, each with fewer carbon atoms than the original
ketone.
Aldehyde and Ketone Tests
Tollen's Silver Mirror Test
Tollens' reagent is an ammoniacial solution of silver ion prepared by dissolving silver oxide in ammonia.
R-C-OH + HO-C-R'
Several simple tests which are based on this difference in reactivity toward oxidizing agents are used to
distinguish between the two classes of carbonyl compounds.
The reagent is reduced to metallic silver by aldehydes which, in turn, are oxidized to the corresponding
acids. Ketones are not oxidized by the reagent.

If the test is carried out with dilute solutions of reagents, in scrupulously clean glassware, the silver
deposits finely in the form of a mirror on the walls of the beaker or test tube. Otherwise, the silver is
deposited as a black precipitate.

Fehling's or Benedict's Test

The reagent in each of these tests is cupric ion, Cu2+ in an alkaline solution. To keep the copper from
precipitating as the hydroxide, it is complexed with either tartrate ion (Fehling's reagent) or citrate ion
(Benedict's reagent). For simplicity's sake we will write the equation with just the cupric ion. Aldehydes
reduce the copper; the solutions usually turn from blue to muddy green, and gradually a reddish
precipitate of cuprous oxide, Cu2O is formed. Most simple ketones do not react, although certain
hydroxyketones and carbohydrates do.

4. Account for the differences in the reaction of starch, sucrose, and their hydrolysates with Benedict’s
reagent.
- Benedict's test uses a chemical which will react with reducing sugars (like glucose and fructose) to
produce a precipitate. Since sucrose and starch are not reducing sugars, Benedict's test will be negative
for them. However, the hydrolysis of sucrose produces glucose and fructose, both reducing sugars. So, if
sucrose has been hydrolyzed, there will be reducing sugars present which will react with Benedict's
reagent to give you a positive test. Similarly, hydrolysis of starch produces glucose which will react with
Benedict's reagent.

5. What is the structural requirement for the reducing property of carbohydrates?

- Sugars may be classified as reducing or non-reducing based on their reactivity with Tollens', Benedict's
or Fehling's reagents. If a sugar is oxidized by these reagents it is called reducing, since the oxidant (Ag(+) or
Cu(+2)) is reduced in the reaction, as evidenced by formation of a silver mirror or precipitation of cuprous
oxide. The Tollens' test is commonly used to detect aldehyde functions; and because of the
facile interconversion of ketoses and aldoses under the basic conditions of this test, ketoses such as
fructose also react and are classified as reducing sugars.
When the aldehyde function of an aldose is oxidized to a carboxylic acid the product is called an aldonic
acid. Because of the 2º hydroxyl functions that are also present in these compounds, a mild oxidizing agent
such as hypobromite must be used for this conversion (equation 1). If both ends of an aldose chain are
oxidized to carboxylic acids the product is called an aldaric acid. By converting an aldose to its
corresponding aldaric acid derivative, the ends of the chain become identical (this could also be
accomplished by reducing the aldehyde to CH2OH, as noted below). Such an operation will disclose any
latent symmetry in the remaining molecule. Thus, ribose, xylose, allose and galactose yield achiral aldaric
acids which are, of course, not optically active. The ribose oxidation is shown in equation 2 below.
Other aldose sugars may give identical chiral aldaric acid products, implying a unique configurational
relationship. The examples of arabinose and lyxose shown in equation 3 above illustrate this result.

6. Give a simple chemical test to differentiate between the following. Show equations for the reactions.
a. butanone and butanal (butyraldehyde) b. 2-propanol and acetone c. glucose and butanal d. sucrose
and lactose e. glucose and 1-pentanol