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Chemistry of life (Biochemistry)
The study of chemical compounds that are vital for
living organisms to sustain life is called biochemistry. The
subject deals with the nature of these compounds and
characteristic reactions they make inside the living
organisms . We are not involved fundamentally with the
study of biochemistry as a subject , but to give brief
introduction to main classes of the organic compounds in
this important field. It is beyond this discussion to present
detailed explanation of these essential organic substances .
We will give short introduction of the main classes and
their active role in our body . Some of these groups are ,
carbohydrates , fats and proteins, etc..
· Carbohydrates.
Carbohydrates are classes of organic compounds that
consist of carbon , hydrogen and oxygen with an empirical
formula of Cm(H2O)n in most cases . The terms m and n
can be the same as in the case of C6H12O6 (glucose) or
different in the case of C12H22O11 (sucrose) . Another
important feature of the carbohydrates is that oxygen and
hydrogen are generally in ratio of 2:1 , so that it was
historically called hydrates of carbon ; but not all
compounds of carbohydrates necessarily maintain this
hydrogen – oxygen ratio and not all compounds that fit
this hydrogen-oxygen ratio are carbohydrates .
In biochemistry the term carbohydrate denotes different
compounds called saccharides . These compounds include
sugars , starch and cellulose . Saccharides (Greek word
meaning sugars) are generally classified into
monosaccharides , disaccharides and polysaccharides .
Monosaccharides are the simple sugars which are either
aldoses (aldehydes) like glucose or ketoses ( ketones) like
fructose . These simple sugars are further classified on the
base of the number of carbon atoms they contain like
pentose (containing five carbon atoms) , or hexose
(containing six carbon atoms) .
Carbohydrates are naturally formed in a process called
photosynthesis in which plants combine CO2 from the air
and water from the soil in the presence of chlorophyll ,
sunlight and certain enzymes producing simple sugars .
6 CO2 + 6H2O (sun light) C6H12O6 + 6O2
sugar(glucose)
This above reaction is not simple process as it looks ,
but extremely complicated reaction with different
intermediate steps before it gives the final product . since
the final product is a monosaccharide , plants have the
ability to synthesize disaccharides by combining two
molecules of monosaccharides .
2 C6H12O6 C12H12O11 + H2O
monosaccharide
disaccharide
This above natural chemical process is the reverse of
hydrolysis . Both plants and animals have the ability to
synthesize polysaccharides by combining large number of
monosaccharide molecules .
n C6H12O6 (C6H10O5)n + n H2O
polysaccharide in plants are usually in the form of
cellulose , stored in stalks and stems and in a form of
starch in the roots and seeds .
Animals and plants depend on each other to sustain life ;
plants can synthesize carbohydrates - as we mentioned
before - from simple inorganic materials like CO2 and
H2O while animals cannot. On the other hand , animals
utilize these essential carbohydrates and metabolically
oxidize to give CO2 , water and energy .
C6H12O6 + 6 O2 6 CO2 + 6H2O + energy
Note that this reaction is the reverse of that of the
photosynthesis in plants ; we can combine both equations
to establish the relationship .
The reaction to the right is endothermic and the energy
is taken from the sunlight while the reaction to the left
is exothermic in which the same energy is liberated and
utilized by the body activity to sustain life .
It is clear from this cycle that the energy stored in the
carbohydrates is originally from the sun .This energy is
used by all living organisms in their metabolic process.
1- Monosaccharides
Monosaccharides are the basic units of the carbohydrates
; they are also called simple sugars . The term
monosaccharide originally came from two Greek words
mono- which means single and Sachar which means sugar .
These simple sugars are usually colorless crystalline solid
material . Almost all monosaccharides are soluble in water
and have sweet taste .
Most monosaccharides have chemical formula of
Cm(H2O)n where n < 2 . They are classified - as we already
described - on the base of number of carbon atoms present
in the molecule such as triose , pentose , and hexose
representing glyceraldehyde , ribose and glucose respectively
.
The carbon chain of the monosaccharides are numbered
beginning from the carbon closest to the carbonyl group .
We can also designate the structural nature of the
monosaccharide by using the terms aldoses or ketoses so
that if the carbonyl is at position 1 of the carbon chain ,
the sugar is an aldehyde and termed an aldose . On the
other hand if the carbonyl group is in between two
carbon atoms , then the sugar is ketone and termed ketose .
· Structures of monosaccharides – (optical isomers).
Monosaccharides exist in different stereo-isomeric forms .
In section (10.3) of this chapter , we presented this
property in detail and explained the two types of the
stereoisomers ; the important one in our study of simple
sugars is the optical isomerism .
Optical isomers are molecules that have the same
molecular formula but differ in the arrangement of the
atoms in space in such a way that they are mirror images
to one another . Despite these isomers have the same
physical and chemical properties , they differ in the
direction of rotating planely polarized light (light waves
vibrating in one plane ) . If polarized light is passed
through these isomers, the plane of the vibrating light is
rotated due to presence of chiral or asymmetric carbon in
the molecule that is a carbon atom attached to four
different groups . Suitable example is the glyceraldehyde
molecule .
The central carbon atom is connected to aldehyde group
, OH group , hydrogen and alcoholic group . Despite
glyceraldehyde is the simplest carbohydrate , it is very
important compound in the since that many complex
sugars can be derived .
· Classification of monosaccharides
We have seen that simple sugars can be divided into
different groups depending on the number of carbon atoms
they contain designating them as trioses , tetroses, pentoses
and hexoses.
A. Trioses: These are monosaccharides that consist of three
carbon atoms . Glyceraldehyde is an example of this group
. Trioses are the product of the metabolic breakdown of
certain simple sugars .
B. Tetroses: Tetrodes are simple sugars with four-carbon
structure . Erythrosine and throes are examples of these
sugars .
C. Pentose: These are monosaccharides with five-carbon
atoms . They are classified into two groups ; those with
aldehyde functional groups called aldopentose and those
with ketone functional groups called ketopentose .
The most significant of these sugars are the ribose and
deoxyribose which are found in RNA (Ribonucleic acid)
and in DNA (Deoxyribonucleic acid) . Both RNA and DNA
are the main component of every living cell , especially in
the nucleus of the cell .
D. Hexoses: They are monosaccharides with six carbon
atoms . There are different kinds of hexoses but the most
important ones in terms of human body are glucose ,
fructose and galactose . These sugars have the same
chemical formula but differ in structure and can be called
isomers of a given molecular formula .
We already explained that simple sugars are classified on
the base of their functional groups . Hexose with aldehyde
functional group are called aldohexose and those with
ketone functional group are called ketohexose .
A. Glucose: glucose is an aldohexose with general formula
C6H12O6 ; there are four asymmetric carbon atoms and 16
optical isomers . Glucose molecules can exist in solution in
the form of open chain or in the form of ring structure .
This ring structure is formed when the hydrogen of the
carbon 5 -OH attaches with the oxygen atom of the
aldehyde group on carbon 1 .
Open chain
Ring structure
B. Galactose: galactose is an aldose sugar like glucose ; it
can be considered as the isomer of glucose , they only
differ in the configuration of a single carbon atom which
is known as epimoric form of isomerism (differ in one
stereoisomeric center) . Like any hexose , it can exist in
solution as an open chain or ring form .
Open chain
Ring structure
C. Fructose: fructose is ketohexose with a molecular
formula of C6H12O6 , like glucose and galactose . Fructose
or levulose is naturally occurring sugar found in fruits ;
like any other hexoses , it is found in solution as a ring
or as open chain structure . Fructose is the sweetest and
most soluble of all sugars .
Reactions of the hexose monosaccharides
Aldoses and ketoses have reducing properties in common
. These properties are the bases of the chemical tests
undertaken to determine the sugar content in human blood
and urine . The reactions involved in these chemical test
are in the organic practical section . In these tests aldoses
are oxidized into their corresponding acids .
RCHO + Cu2+ + 2 OH-
RCOOH + Cu2O + H2O
Clinitest tablets are used to perform the above test to
determine the sugar in the human urine . Note that these
tablets contain copper ions in the form of Cu(II)sulfate.
Another test for reducing sugars is the Tollens test in a
solution containing silver ions in basic solution .
Glucose + Ag+ + OH- gluconic acid + Ag +
water
Another general qualitative test for carbohydrates is the
Molisch test in which concentrated sulfuric acid is
carefully added to the testing solution containing alpha-
naphthol ; formation of purple ring at the interface of the
acid and solution confirms the presence of carbohydrate
substance .
Another important reduction reaction for some aldoses like
glucose is the fermentation process in which an enzyme
from the yeast catalyzes this reaction and without which
this reaction cannot take place .
C6H12O6 enzyme CH3CH2OH + CO2
Aldoses can also undergo oxidation since they both
contain aldehyde and hydroxyl group . We have seen that
oxidation of the aldehyde group gives the corresponding
acid , like gluconic acid in the case of glucose and
glucuronic acid if the hydroxyl group oxidizes .
Aldohexoses can be converted to alcohols by reduction and
in this case , glucose is reduced to sorbitol and fructose is
reduced to mixture of mannitol and sorbitol.
Glucose
Sorbitol
Since both sorbitol and mannitol are partially and slowly
taken by the body , it is useful as sweetening agent for
the diabetic people . When orally taken some of the sugar
is excreted in the urine and other is taken by the body
slowly .
1. Disaccharides
Disaccharides are the sugars formed when two
monosaccharides are chemically joined together . The most
common examples of the disaccharides are sucrose , lactose
and maltose . They all have the general formula
C12H22O11 , which means they are isomers .
Disaccharides formed as a result of condensation reaction
of two monosaccharides which means elimination of water
molecule . The reverse reaction is known as hydrolysis
performed by certain enzyme .
These reactions are vital in the metabolism of living
things . Different disaccharides hydrolyze to give two
different or same simple sugars as shown above.
Disaccharides are , like monosaccharides , white crystalline
sweet solids . Their solubility varies from completely
soluble to slightly soluble .
Disaccharide molecule from two simple sugars is formed
by displacing hydroxyl group from one molecule and the
hydrogen ion from the other producing water molecule a
process known as condensation reaction .
Sucrose is formed by the reaction of the aldehyde group
of glucose with the ketone group of fructose thus
eliminating the reducing property of the carbonyl; and for
this reason give negative test to reducing reagents. On the
other hand , maltose and lactose have free aldehyde group
and give positive test for both Fehling and Tollens
reagents .
In the case of maltose formation , one aldehyde group of
a glucose reacts with the hydroxyl group of another
glucose leaving one aldehyde group free for reducing
property . In the same way , lactose has one available
aldehyde group ; for that reason maltose and lactose are
reducing sugars while sucrose is not reducing sugar .
These sugars are differentiated analytically on the base of
their fermentation and reducing properties . Both sucrose
and maltose can be fermented by the addition of yeast
which contains sucrase and maltase enzymes . Lactose does
not ferment with the yeast since it does not contain
lactase .
A. Sucrose:
This sugar is known as table sugar ; it is naturally
occurring carbohydrate found in many fruits and vegetables
like carrot roots and pineapples . Sucrose is extracted from
cane and beet sugars and refined to be consumed by the
people. Hydrolysis of sucrose gives a mixture of glucose
and fructose .
This mixture of equal amounts of glucose and fructose is
invert sugar . High concentration of this invert sugar is
found in honey ; this kind of sugar is less prone to
crystallization when cooled .
B. Maltose:
Maltose is a disaccharide formed from two units of
glucose . It is commonly known as malt sugar . It is
commercially prepared from hydrolysis of starch . It
generally occurs in germinating seeds .
C. Lactose:
Lactose is also a disaccharide formed from joining of
two units of glucose and galactose . It is found in milk
and that is the reason why it is called milk sugar ; it
differs from the above sugars in the sense that it comes
from animals. We pointed out previously that lactose is
fermented by the enzyme lactase forming lactic acid; this
fermented milk is said to be sour milk. Lactose is also
found in urine of pregnant woman and since it is
reducing sugar , it gives positive test with Fehling or
Benedict solution.
2. Polysaccharides:
Polysaccharides can be defined as the polymeric form of
monosaccharides . These polymers hydrolyze to yield many
monosaccharide molecules . These polysaccharides are
generally formed from five-carbon or six-carbon sugars .
The most common polysaccharides are :
A. Starch
Starch is a polymer of glucose sugars which consist
of around 20% amylose and 80% amylopectin . Amylose is
linear chain of glucose molecules while amylopectin is
branched form of the starch and contain much larger units
of glucose in comparison to amylose . Starches are
insoluble in water .
Starch (long chain of glucose units )
Amylopectin (branched form of starch)
Amylose (linear chain of glucose)
The presence of starch is tested with iodine which gives
characteristic deep blue color . Hydrolysis of starch to
simple sugars through several stages giving different
dextrin’s on the process which can be followed by
addition of iodine . Iodine gives no color with simple
sugars and blue color with dextrin’s .
A. Cellulose
Cellulose is also polysaccharide consisting of linear
chain of several hundred to thousands of linked glucose
units . It is an important supporting and structural
component of all plants . Unlike starch , cellulose is not
affected by the digestive human enzymes and for that
reason helps prevent constipation by making bulk feces of
dietary fiber when taken by human .
Cellulose
Cellulose is digested and metabolized by certain animals
like ruminants and termites with the help of micro-
organisms which live in their abdomen . Cellulose is not
soluble in water and in majority of solvents . Unlike starch
, it does not give positive test to iodine or to Fehling
reagent . Materials like cotton , wood and paper are mainly
composed of cellulose . Cellulose is used to produce paper
, cellophane and rayon.
B. Glycogen ,
Glycogen is multi-branched glucose polymer which is
responsible for the storage of energy in animals . Glycogen
is produced by the body and stored in the liver and the
muscles ; it is originated from the animals while starch is
basically from plant origin ; it acts as a reserve or long-
term energy storage . Even though fats act as the primary
energy stores , glycogen energy is necessary for immediate
mobilization to cover a sudden need for glucose.
Glycogen is synthesized in the body cell from glucose
through a process called glycogenesis and hydrolyzed to
give glucose through a process known as glycogenolysis .
Glucose glycogenesis
Glycogen glycogenolysis Glucose
C. Dextrin
Dextrin’s are lower molecular weight carbohydrates formed
as a result of starch hydrolysis . Dextrin’s are considered
as an intermediate between the starch and the
disaccharides . It dissolves in water giving sticky colloidal
suspension used in the preparation of adhesives like the
glue on the back of the postage stamp .
1. Lipids (fats)
Lipids are naturally occurring organic compounds ; these
include fats , waxes , fat soluble vitamins and sterols
(steroid alcohols) . It is one of the main sources of food
essential for animals and plants .
Lipids consist of carbon , hydrogen and oxygen and in
certain cases nitrogen and phosphorus . They are insoluble
in water but soluble in organic solvents like alcohol and
acetone . Even though there are different categories of
lipids , we will confine our discussion to simple lipids
which are esters of fatty acids .
· Fatty acids:
There are two types of fatty acids ; saturated fatty acids
which contain single bond between the carbon atoms and
unsaturated fatty acids which contain double between the
carbon atoms .
CH3-CH2-CH2-CH2-COOH CH3-(CH2)7-
CH=CH-(CH2)7-COOH
Saturated fatty acid
unsaturated fatty acids
Table.10.6 Common saturated fatty acids.
Name
Formula
Source
Butyric acid
C3H7COOH
Butter fat
Caproic acid
C5H11COOH
Goat fat
Caprylic acid
C7H15COOH
Coconut oil
Capric acid
C9H19COOH
Palm oil
Lauric acid
C11H23COOH
Laurel (plant)
Table.10.7 Common unsaturated fatty acids .
Name
Formula
Source
Oleic acid
C17H33COOH (1 double bond)
Olive oil
Linoleic oil
C17H31COOH (2 double bond)
Linseed oil
Linolenic acid
C17H29COOH (3 double bond)
Linseed oil
Unsaturated fatty acids can exist in two structural forms
, cis in which the two hydrogen atoms adjacent to the
double bond are on the same side of the carbon chain
and trans in which the adjacent two hydrogen atoms are
on the opposite sides of the carbon atom (discussed in
section 10.3) .
Iodine number
Human body cannot sufficiently synthesize some of the
fatty acids they need and must be obtained from the food
they take . These fatty acids are known as essential fatty
acids . linoleic and linolenic acids are among the essential
acids ; they are found in certain plant oils like peanut ,
soybeans and corn but not in olive oil or cocoanut.
Essential fatty acids have important function in the body
such as the synthesis of prostaglandins (hormone-like
substance with wide physiological activities) .
Iodine number:
The degree or the amount of unsaturation in a given
fatty acid is determined by its iodine number , which is
the amount of iodine consumed by 100 g of the acid.
Iodine reacts with the double bond of the unsaturated
fatty acid , so the higher the iodine consumed the more
double bonds present in that fatty acid .
Despite there are different methods for determining
iodine value , they all fall under the iodiometry procedures
. We indicated that the higher the unsaturation for a fatty
acid the higher the iodine value . Linseed oil has much
more iodine value than coconut oil which means the
linseed oil is highly unsaturated oil with respect to
coconut oil . In general , vegetables oils have higher iodine
number than animal fats .
Fats and oils
We defined the fats as esters of fatty acids (already
discussed in carboxylic acids unit) which are formed from
the reactions between a fatty acid and specific alcohol
called glycerol .
There are three OH groups available in glycerol and
each one can react with a fatty acid molecule forming a
fat molecule . These fatty acids can be of same fatty acids
or different fatty acid molecules .
The esters formed when the alcohol is glycerol is called
glyceride . It can be monoglyceride , diglyceride or
triglyceride depending on the number of fatty acids
reacted with the glycerol .
In general if the reacting fatty acid is saturated , the ester
formed is called fat and if it is unsaturated the resulting
ester will be an oil . Most fats have iodine value less than
70 while most oils have iodine value more than 70 . Keep
in mind that mineral oil is a saturated hydrocarbon
different than fats and oils which are either from animal ,
vegetable origin and petroleum distillate .
Those oils are different from essential oils which are
volatile liquids used as flavors and perfumes , while
mineral oils are mixtures of higher alkanes . The name
may be inappropriate and sometimes other names are
applied such as liquid paraffin , paraffin oil etc. .
· Properties and uses of fats
Fats can be considered as body fuel , they produce
more energy for the body than any other nutrient such as
carbohydrates or proteins . Another important function of
the fats is their essential role in reserving food supply for
the body . They protect the vital organs by keeping them
rigid in their place and helping to absorb shocks .
In terms of physical properties , fats and oils are either
in liquid or solid state with white or yellow color in
most cases . They are colorless and tasteless in their pure
state but get rancid as the time goes on creating
unpleasant odor and taste. Since both fats and oils are
insoluble in water ; they form transient emulsions which
can be stabilized by adding emulsifiers from the bile to
make them digestible .
In terms of chemical properties , fats hydrolyze in the
presence of certain enzymes to form fatty acids and
glycerol . We have seen that when a fat is formed water
is the product (see the preceded equation) ; the reverse
process is called hydrolysis.
After the hydrolysis of the fat , the glycerol is separated
and purified for medical and industrial purposes . It is
used as humectant (against loss of moisture) for skin and
hair care products . It is also used in the preparation of
medical and pharmaceutical products . Glycerol has also an
important industrial applications ; it is found in food ,
beverages and sweetener as sugar substitute besides many
other uses .
We briefly explained in this chapter , that basic
hydrolysis of fats yield glycerol and salt of the fatty acid
; this process is called saponification and the sodium or
potassium salt formed is called soap .
Sodium soaps are solid bars and potassium soaps are
liquid . In preparing soaps, various materials are added to
give pleasant odor and color ; in certain soaps , germicidal
substances are added to protect the skin from the germs
as in the case scouring soap .
In hard water , calcium and magnesium ion react with the
soap rendering it insoluble and inactive to give more
lather and more cleansing result . This problem is
overcome by using detergents . Detergents are synthetic
cleaning substances consisting of mixtures of surfactants
(lower the surface tension) used as a soap . They can be
used both in soft and hard water ; which means calcium
and magnesium salts of detergents are soluble in water
thus overcoming the disadvantage of soap .
Detergents are made from different petrochemicals besides
fat and oils . They are generally neutral unlike the soaps
which are alkaline . The sodium salt of the detergents is
prepared by the reaction of long chain alcohols with
concentrated sulfuric acid and the product is neutralized
by sodium hydroxide .
R-OH + H2SO4 R- OSO3H + H2O
long chain alcohol
R- OSO3H + NaOH R-OSO3Na + H2O
Detergent
The unbranched straight chain detergents are less harmful
in terms of environmental pollution .
2. Proteins
Another essential nutrient for living organisms is the
protein . Proteins are complex organic substance which
consist of carbon , hydrogen , oxygen and nitrogen besides
some other elements like sulfur and phosphorus . Proteins
consist of long chain of simple molecules called amino
acids arranged in a specific order. There are 20 known
amino acids .
All of these amino acids cannot be synthesized by the
human body and must be externally supplied . The first
three amino acids are shown below with their symbol and
names
Amino acid molecule contains amino group and carboxylic
group .
Despite the amino group can be structurally anywhere in
the molecule , the amino group of the naturally occurring
amino acids are bonded with the alpha – carbon (the one
next to acid group ). Almost all amino acids have alpha
chiral carbon (except glycine) and show L-configuration .
Amphoteric nature of the amino acids
Amphoteric substance is one that shows both acidic and
basic character ; which means it behaves as an acid and
base. As the name denotes , amino acids contain carboxylic
group which is an acid since it furnishes proton in
solution and also contain amino group which accepts the
proton and on the base of Brnsted – Lowry acid-base
definition , amino acids act as an acid and as a base at
the same time; therefore amino acids react with both
acidic and basic substances ; consequently amino acids can
be called amphoteric substance as long as they donate and
accept protons .
Amino acid molecules react together in which the amino
group of one amino acid reacts with the carboxylic group
of another forming a long chain of amino acid molecules
connected through a bond known as peptide bond . This
bond is formed as a result of condensation reaction in
which one molecule of water is produced for every two
reacting amino acids .
A combination between two amino acids form a product
called dipeptide , and that of three amino acids form
tripeptide and more than three amino acids form
polypeptide. There are two different ways for two amino
acids to combine .
It is very important to know that for combined amino
acids , their name is abbreviated by taking the first three
letters of the amino acid and in the order of their
combination in the peptide ; the first amino acid is the
one that furnishes the OH from the acid group .
· Isoelectric point
We have seen that amino acids contain both basic and
acidic functional groups. In the crystalline forms , amino
acids exist as zwitterions , which is a neutral molecule
having both positive and negative electrical charges
resulting no net electric charge. These solid forms usually
have high melting point and less solubility in organic
solvents but soluble in water . In aqueous solution , these
amino acids establish an equilibrium dependent on the pH
of the solution .
isoelectric
point
In basic solution , the equilibrium is shifted towards the
right and in acidic solution the equilibrium is shifted
towards the left . If two electrodes are placed in this
amino acid solution , there will be net migration of the
solute molecules towards either the positive electrode
(cathode) or the negative electrode (anode) depending on
the alkalinity or acidity of the solution .
In basic solution a negatively charged carboxylic group is
formed and the migration takes place towards the
positively charged electrode and in acidic solution , a
positively charged amino group will be formed and the
migration takes place towards the negatively charged
electrode .
At certain pH , characteristic of each amino acid , the
positive and the negative ions will be equal and there
will be no net migration of the amino acid molecules
towards the electrodes . This specific pH value is called
isoelectric point .
Proteins like amino acids have an isoelectric point which
is characteristic property for each protein . Proteins have
the minimum solubility in their isoelectric point .
For the pH of the amino acid above the isoelectric point ,
the basicity dominates , that is there will be more basic
sites and the pH below the isoelectric point , the acidity
will dominate and there will be more acidic sites .
· Structure and the properties
Proteins hydrolyze in acidic or basic media , or by the
action of certain enzymes . This hydrolysis breaks the
protein molecule into the constituent amino acids . In the
reverse process , amino acids combine together with the
help of specific enzymes to form polypeptides which are
the basic molecular structures of the proteins ; the more
units of amino acids combined the bigger the molecular
weight of the protein .
As a result of the large molecular sizes of the proteins ,
different structural forms are created which are referred to
as primary , secondary and tertiary structures based on the
shape and spatial orientation of the protein molecule .
The bases of these shapes and their biological significances
is beyond our discussion. The main properties of the
proteins can be summarized as follows :
A. Colloidal
The aqueous solution of protein form colloidal dispersion
of the protein molecules in water . These colloidal particles
have the ability to pass through the filter paper but inable
to pass thhrough the cellular membranes ; this phenomenon
is physiologically important since it keeps the protein in
the bloodstream and impedes the diffusion of protein
molecules into the urine indicating the damage of kidney
membranes .
B. Denaturation
Denaturation refers to change of protein structure loosing
its biological function . When proteins become denaturated ,
they coagulate or precipitate . This effect takes place under
the influence of several reasons :
1. Effect of alcohol: 70% aqueous solution of alcohol
(ethanol) precipitates almost all proteins and for this
reason we use this alcohol concentration as a disinfectant
because it coagulates the protein of the bacteria rendering
it inactive .
2. Salting out process: at high concentration of salt solution
, proteins become almost insoluble and precipitate . This is
one of the techniques used to separate proteins from
solutions . Salts like Na2SO4 , NaCl are used for this
purpose . The separation from the solution is achieved
through filteration and the traces of the remaining salt is
eliminated by a process known as dialysis .
3. Heavy metal salt : Saturated salt solutions of certain
heavy metals such as silver and mercury(II) precipitate the
protein . These heavy metal salts coagulate the protein
making it biologically inactive . This is the reason why
these metals are poisonous if taken internally .
4. Heating :
Almost all protein coagulate on heating . Heating is
common method to sterilize materials because the protein
of the bacteria is destroyed by coagulating it . Presence of
protein in urine (which is kidney malfunction) can be
determined by heating given sample of urine to
precipitate the protein .
5. Concentrated inorganic acids:
Concentrated strong mineral acids can be used to
precipitate the protein. HCl , H2SO4 , and nitric acid are
among the mineral acids that can be employed to
precipitate proteins . Concentrated HCl from the gastric
glands precipitate the milk protein , casein , in the form of
curds .
C. Qualitative tests for proteins
Most qualitative tests for proteins depend on specific color
formation . These tests are not always decisive since it
depends on the presence of certain amino acids . Some of
these tests are :
Xanthoprotic : Which is the formation of yellow precpitate
due to action of nitric acid on proteins . Contact of nitric
acid on your finger usually produces yellow color on the
skin of the finger . This test is only positive for the
amino acids that contain benzene ring .
There are other well known tests like , Biuret , Millon’s ,
Hopkin-Cole and ninhydrin tests in which their procedures
are explained in the organic practical section. Different
chromatographic methods are used to separate and identify
different mixture of proteins or amino acids .
Quantatively the amount of protein in a sample is
estimated by determining the nitrogen content and
multiplying the result by factor 6. This factor came from
the fact that the average percent of nitrogen in protein is
16% by weight ; that is approximately 1/6 of the total
weight of the protein . If for example the total nitrogen of
a sample is 2.5 g , the amount of protein present in the
sample = . The result is always expressed in percent .
Nucleoproteins
Nucleoproteins can be defined as conjugated propteins that
consist of acidic part which is nonprptein section called
nucleicacid and a protein part . Nucleoproteins hydrolyze
during the digestion process into nucleic acids and
proteins . Proteins further hydrolyze into amino acids while
nucleic acids hydrolyze into nucleotides .
Nucleic acids are biological polymers that consist of
repeating units of nucleotides so that nucleic acids can be
called polynucleotides . Each nucleotide consist of
heterocyclic nitrogen base , pentose sugar and phosphoric
acid . The number of nucleotide units in a nucleolic acid
depends on the nature of the nucleic acid and may range
from approximately one hundred units to several million
units .
Nucleotide
There are mainly two branches of nucleic acids :
a) Deoxyribonucleic acid (DNA) which is found in the
nucleus of the cell and contains pentose sugar ,
deoxyribose , which is the only sugar on hydrolysis.
b) Ribonucleic acid (RNA) which is mainly found in
cytoplasm and contain ribose as the only sugar on
hydrlysis .
Parts of nucleotides
We have showed that a nucleotide consist of nitrogen base
, pentose sugar (ribose or deoxyribose ) and phosphoric acid
. Let us investigate them one by one .
A. Nitrogen bases:
The two groups of heterocyclic nitrogen – containing bases
in the nucleic acids are derivatives of purine or
pyrimidine with following structures .
The most common purine bases in nucleic acids are
adenine and quinine . They are present in both DNA and
RNA .
The pyrimidines that are most common in both DNA and
RNA are cytosine , uracil and thymine .
(2-oxy-6-aminopyrimidine) (2,4-dioxy-5-
methylpyrimdine) (2,dioxypyrimidine)
Nucleosides ,
Nucleosides are units formed by combination
of nitrogenous base and one of the two pentose sugars
(ribose or deoxyribose) in the structure of nucleic acids .
In this combination the glycosidic bond between the sugar
and the nitrogenous base is formed at C – 1 of the sugar
and the position 9 of the purine base or position 1 of the
pyrimidine base .
Glyosidic between C1 of ribose
Glyosidic between C1 of deoxyribose and
and positon-9 of purine
postion-1 of pyrimidine
Nomenclature
Nucleosides from the pyrimidines have names ending in -
idine and those from the purines have names ending in -
osine Purines with ribose sugars are named as follows :
Adenine + ribose Adenosine
Guanine + ribose Guanosine
Purines with deoxyribose sugars are named as follows :
Adenine + deoxyribose
Deoxyadenosine
Guanine + deoxyribose Deoxy
guanosine
Pyrimidines containing ribose sugars are named as follows
:
Uracil + ribose Uridine
Cytosine + ribose Cytidine
Thymine + ribose Thymidine
Pyrimidines containing deoxyribose sugars are named as :
Uracil + deoxyribose Deoxy uridine
Cytosine + deoxyribose Deoxycytidine
Thymine + deoxyribose Deoxythymidine
It is known that glycosides are quite stable in alkaline
medium and the situation is the same in the nucleosides
but hydrolyze in aqueous acid solution to produce base
and pentose sugar .
Polynucleotides Hydrolyze Nucleotides Hydrolyze
Nucleosides Hydrolyze Base + sugar
Nucleotides are the phosphate esters of the nucleosides .
The phosphoric acid reacts with one of the hydroxyl
groups in the sugar forming phosphate ester bond.
Condensation reaction
In the case of ribonucleoside , the phosphate ester bond
can be formed at three possible -OH groups at C1 , C3
and C5 and for that reason we have to specify the point
of the attachment in naming the nucleotide .
We already pointed out that nucleotides are phosphate
esters of nucleosides ; and because the compound contains
phosphoric acid , acidic name is given according to the
following table.
Nucleoside + H3PO4
Acidic name of the nucleotide
Adenosine
Adeynilic acid
Guanosine
Guanilic acid
Cytidine
Cytidilic acid
Uridine
Uridylic acid
Thymidine
Thymidilic acidic
The nucleoside formed from the reaction of one molecule
of a nucleoside with one molecule of a phosphoric acid is
called monophosphate of that nucleoside . In the case of
adenosine , it is called adenosine monophosphate (AMP) or
Adeynilic acid . If it reacts with one more phosphoric acid
molecule , it is called adenosine diphosphate (ADP) ; and if
three groups are involved it is called adenosine
triphosphate (ATP) .
) AMP)
(ADP)
(ATP)
These phosphate esters , AMP , ADP and ATP perform
vital role in cellular metabolism as high storage of energy
and release that energy from the hydrolysis of phosphate
bonds and pass to the specific reactions that require this
energy to take place . ATP contains the highest amount of
energy in comparison to ADP and AMP. Besides being
body building blocks , certain nucleotides perform
biological activities of their own . Appropriate example can
be taken in the case of cyclic adenosine monophosphate
(cyclic – AMP) which is formed as a result of phosphate
group attached to carbon-3 and carbon-5 of the ribose
sugar . It (cyclic-AMP) acts as messenger in regulating
enzymatic activities in cells that store fats and
carbohydrates.
Cyclic adenosine monophosphate (camp)
Certain nucleotides act as Coenzyme . Nicotinamide
adenine dinucleotide (NAD+) is essential for many
biological oxidation-reductions reactions to take place . It
exists in oxidized form (NAD+) and reduced form
transferring electrons from reaction to another .
Exercises:
1. What is the modern definition of organic chemistry ?
Briefly explain the role of organic chemistry in our daily
life.
2. What was the vital force theory and how was it
disproved ?
3. Account for the fact that element carbon has the unique
property of forming so many millions of chemical
compounds .
4. Briefly compare , in general , the physical and chemical
properties of organic and inorganic compounds .
5. Explain the role of carbon dioxide in the natural
formation of certain organic substances like coal ,
petroleum , and natural gas .
6. What are the hydrocarbons ? mention the different classes
of the hydrocarbons on the base of their general structure
. Give an example of each class .
7. Name the following hydrocarbon compounds .
8. Write the structural formula for each of the following
compounds .
A. 3-Methylheptane
B. 2,3-Dimethylpentane
C. 2-Chloro-3-ethylpentane
D. 1,1,2,2-Tetrabromobutane
E. e) 2-Butyne
F. 3,4-Diethyl-1-hexyne
G. 3-Ethyl-2-heptene
9. Write the possible isomers for the following
hydrocarbons .
A. C7H16
B. C6H12
C. C5H10
D. C4H8
E. C6H10
10. How can you determine the nature of the following
hydrocarbons as alkanes, alkenes, alkynes and cycloalkanes .
A. C7H14
B. C6H12
C. C5H12
D. C4H6
E. C3H4
11. How can you distinguish by simple qualitative test
between the following pairs of compounds ?
A. Ethanol and pentane .
B. Acetaldehyde and acetone .
C. Acetic acid and ethylamine .
D. Phenol and benzene .
E. Pentane and pentene .
12. Account for the extra stability of benzene in
comparison to ethene (ethylene).
13. Explain what we mean by a functional group . Why it
is important to classify organic compounds on the base of
their functional group ?
14. Distinguish the following compounds as alcohol ,
aldehyde , ketone , carboxylic acid and amine .
A. CH3-COCH3
B. CH3CH(OH)CH3
C. HCHO
D. CH3CH2CO2H
E. CH3CH(NH2)CH3 .
15. Draw the possible structural formula of the following
compounds .
A. C2H6O
B. CH4O
C. C3H6O2
D. C3H8O
E. C2H7N.
16. Complete and balance the following equations .
17. Define the following terms .
A. Fatty acids
B. Esters
C. Oils
D. Acid chlorides
18. Give an example of each of the following reactions .
A. Addition reaction
B. Substitution reaction
C. Elimination reaction
19. What are nucleoproteins ?
20. Explain briefly the hydrolysis product of nucleic acid
.
21. What is the basic difference between the RNA and DNA
in terms of structure ?
22. Write T at the end of the true statement and F at the
end of the false one.
A. Adenosine consist of ribose sugar and pyrimidine base .
B. Hydrolysis of nucleotide gives phosphoric acid and
nucleoside .
C. The nucleoside urosine is made up of uracil and ribose
.
D. The nitrogen containing bases in the nucleic acids are
either derivatives of pyrimidine or pyridine .
E. Uracil is found in both RNA and DNA .
23. Explain the relationship between the plants and
animals in terms of synthesis and utilization of
carbohydrates .
24. Despite that lactose and sucrose have the same
formula (C12H22O11), they give different results to
reducing agents . Explain why ?
25. Write (√ ) at the end of the correct statement and (X)
at the of the incorrect one .
A. Carbohydrates were historically known as hydrates of
carbon . ( )
B. Carbohydrates are synthesized by plants and animals
from inorganic materials . ( )
C. Proteins are essential for living organisms as source of
energy only .
D. Alkaline hydrolysis of fats gives fatty acids .
E. Proteins are polymeric units of amino acids .
F. Large value of iodine number for fats shows more
unsaturated sites of that fat .
G. All the compounds in which the ratio of C to H is 2:1
are necessarily carbohydrates .
H. Nucleosides do not contain phosphate units.
26. Explain the following terms :
a) Fatty acids
b) Isoelectric point
c) Saturated fatty acids
d) Detergents
e) peptide bond.
27. Give reasons for the following statements.
A. All the naturally occurring amino acids are optically active
except glycine .
B. 70% aqueous ethanol is better disinfectant than 100%
ethanol (absolute) .
C. Chiral compounds are optically classified as D and L
forms in their aqueous solutions .
D. Pentose and hexose sugar molecules can exist as ring
form structure .
E. Enantiomers are different than epimers .
C
C
CH
2
OH
OH
OHH
C
C
CH
2
OH
OH
HOH
D -glyceraldehydeL-glyceraldehyde
CHO
C
C
CH
2
OH
OHH
OHH
CH
2
OH
C
C
CH
2
OH
O
OHH
ErythroseErythrulose
CHO
C
C
C
CH
2
OH
OH
OH
H
H
H
H
RiboseDeoxyribose
CHO
C
C
C
CH
2
OH
OH
OH
OH
H
H
H
C
C
C
C
C
CH
2
OH
OH
OHH
HOH
OH
OHH
H
CHO
C
C
C
C
CH
2
OH
HOH
OH
OHH
H
C
C
C
C
C
CH
2
OH
OH
OHH
HOH
H
OHH
OH
O
D-(+)-Glucose
D-(-)-Fructose
D-(+)-Galactose
C
C
C
C
C
CH
2
OH
OH
OHH
HOH
OH
OHH
H
D-(+)-Glucose
CHO
C
C
C
C
CH
2
OH
HOH
OH
OHH
H
O
D-(-)-Fructose
H
2
C
CH
H
2
C
OH
OH
OH
+
RCOOH
RCOOH
RCOOH
H
2
C
HC
H
2
C
O
CH
R
O
OC
R
O
OCR
O
+
3H
2
O
glycerol
fatty acids
triglyceride
Fatty acid+alcohol
Fat+H
2
O
esterification
Hydrolysis
H
2
N - C - H
COOH
H
H
2
N - C - H
CH
3
COOH
H
2
N - C - H
CH
2
OH
COOH
Glycine
Alanine
Serine
H
2
N - C - COOH
R
H
Amino group
Carboxylic group
HOOC - C - NH
2
H
H
+HCl
HOOC - C - NH
3
+
+Cl
-
H
H
NH
2
- C- COOH + NaOH
NH
2
- C- COO- + Na
+
+ H
2
O
H
H
H
H
reactas base
reactas an acid
H
2
N - CH - C
R
1
O
NH - CH - COOH
R
2
Peptide bond
H
2
N - CH
2
- C
O
OH
+ H
2
N - CH - C
O
OH
NH
2
- CH
2
- C -NH - CH - COOH
O
CH
3
Glycine
Gly-ala
+ H
2
O
CH
3
Alanine
O
OH
H
2
N - CH
2
- C
O
OH
Glycine
NH
2
- CH - C - NH - CH
2
- COOH
CH
3
O
H
2
N - CH - CH3
+
Alanine
Ala-gly
H
2
O
+
CH
3
6 CO
2
+ 6H
2
O + ENERGY
plant photosynthesis.
animal metabolism
C
6
H
12
O
6
+6O
2
CH
3
CH
3
Br
CH
3
- CH
2
- CH - CH - CH
3
A-
B-
C-
CH
3
COOH + CH
3
CH
2
OH
C
3
H
8
+ O
2
CH
3
CH
2
OH
[O]
[O]
C
6
H
5
OH + NaOH
C
C
C
C
C
CH
2
OH
OH
OHH
HOH
OH
OHH
H
CH
2
O
C
C
C
C
CH
2
OH
HOH
OH
OHH
H
C
C
C
C
C
CH
2
OH
OH
OHH
HOH
H
OHH
OH
O
D-(+)-Glucose
D-(-)-Fructose
D-(+)-Galactose
Practical Section
A. Wet chemistry practical’s (inorganic portion)
In the following practical work , we will be confined to
experiments based on wet chemistry or bench chemical
analysis ; these are traditional laboratory tests which
encompasses certain processes like sampling , weighing ,
preparing solutions and performing classical chemical tests
. In ordinary wet chemistry analysis , qualitative and
quantitative results can be obtained , yet it is rarely
employed in areas like industry , research centers , forensic
analysis etc. These institutions employ modern chemical
analysis which is extension of the classical analytical
techniques that has been automated and computerized to
save time and increase the sensitivity selectivity of the
system .
The following laboratory sessions involves certain
quantitative topics like gravimetry , titrimetry and
qualitative topics on identification methods based on
formation of color , precipitate or evolution of specific gas
.
Practical chemistry is mere laboratory work in which the
theoretical knowledge of the subject is developed and the
necessary skills are acquired . To undertake this work , the
necessary apparatus , equipment and reagents must be
available and become familiar with the students . For any
given experiment , these materials must be planned and
prepared so that the result can be reported in a scientific
manner . The students must have knowledge of the
application of these tools and get acquainted with the
laboratory environment .
We can classify the laboratory tools into two parts:
A. Glassware: which are glass containers used for mixing ,
measuring , holding and heating laboratory reagents ; they
are generally made from durable and chemically inert
materials like borosilicate’s . These glass containers are of
different shapes and sizes used for different purposes.
Some of these laboratory glassware are stated below .
1. Beakers : Beakers are glass containers of almost
cylindrical shape used for heating mixing and holding
samples . They are available in different sizes .
Volumetric Flaska
Elenmeyer Flaska
2. Flasks : Flasks are narrow-necked and wider bottom
glass containers used for measuring preparing and heating
solutions . Volumetric flasks are used for preparing and
diluting solutions of definite concentrations . Erlenmeyer
flasks or conical flasks are used for heating or handling
solutions .
Boiling Flasks
3. Boiling flasks: flat bottom flasks used for heavy duty
boiling. They are available in different forms and shapes
according to their application.
Test tubes
Different boiling flasks
Test tubes holders
4. Test tubes: Small glass tubes resistant to high
temperature used for testing samples especially
qualitative tests .
Beakers
5. Funnels : Funnels are laboratory apparatus with a wide
mouth and long pipe like opening at the end used to
transfer liquid substance or finely grinded particles to
avoid spillage . They are usually made of glass , plastic or
ceramic .
Different funnels
6. Burettes: Burettes are long graduated glass tubes with
stopcock at end for drawing accurate amount of a solution
. It is extensively used in analytical chemistry, there are
different sizes and structures
Different burettes
7. Other laboratory glassware: Pipettes are used for taking
fixed volume of a liquid. They are usually made of glass or
plastic materials.
Uni-Volume Pipette
Graduated Pipettes
Pipette fillers
Crucibles wash bottles
Desiccator Glass rods
Burette clamp Burette stand Test tube rack
B. Instruments: The other feature of the laboratory equipment
are the instruments. Instruments are devices that help or
facilitate to undertake experimental work .
meter hotplate electronic balance
Mortar and pestle Bunsen burners
Oven Shaker
Gravimetric analysis
One of the most accurate and precise methods in
chemical analysis is the gravimetric analysis . In this
method , the analyte is converted to insoluble form by
precipitating from the solution . The precipitate is then
separated , washed , dried or ignited , weighed and
calculated . Perfect example can be taken in the
determination of Cl- content in a given solution by
precipitating as AgCl with silver nitrate solution . After
filtering and drying , the precipitate is weighed and the
chloride content is calculated .
There are different forms of gravimetric analysis , and the
one we are concerned in this discussion is the one in
which the sought for substance is almost completely
precipitated from a solution so that no considerable
amount is lost during filtration , washing and weighting .To
achieve successful analytical result , the following points
has to be fulfilled :
A. The precipitated substance must be sufficiently insoluble
so that we can analytically ignore the dissolved amount .
B. The precipitate particles must be large enough to be
quantitatively filtered and the impurities can easily be
washed .
C. The final precipitate must be a substance of definite chemical
composition after physical or chemical treatments.
· To accomplish these points and obtain almost fine precipitate
particles, the following problems must be overcome too.
1. Co-precipitation, the precipitate formed may contain foreign
impurities from the solution which depends on the nature of
the precipitate and the conditions in which the precipitate
was formed ; this contamination can be due to either the
adsorption of impurities on the surface of the precipitated
particles in the solution or the occlusion of foreign
particles during the process of crystallization .
2. Coagulation, is the tendency of adsorption layers of the
precipitate to attract ions of opposite charge forming large
size particles containing solvent molecules .
3. Occlusion, this is the way in which foreign particles are
trapped inside the precipitate during the crystallization . It
is not easy to get rid of the occluded impurities .
4. Post-precipitation, it is the precipitation that takes place
on the surface of the precipitating reagent , usually it
comes from a substance that has common ion with
primary precipitate , e. g CaC2O4 / Mg++ & C2O4- - .
Treatment of the Precipitate
To prepare relatively fine pure precipitate , the following
favorable conditions has to be fulfilled.
A. The precipitation must be implemented in dilute solution
keeping in mind the solubility of the precipitate and
digestion period , to minimize the co-precipitation.
B. Add the reagents together slowly with constant stirring
to create large crystals.
C. Precipitation is done in hot solution provided no
counter effect in terms of solubility and the stability of
the precipitate is resulted.
D. The precipitate should be washed with a suitable
solvent.
Precipitation from Homogenous solution
We have already seen that to create a desirable
precipitate , we must minimize the degree of supersaturation
so that dilute solution of the precipitating agent is gently
and slowly added to the sample solution with a constantly
effective stirring . Even though these techniques maintain a low
degree of supersaturation yet excess of the precipitating
reagent is unavoidable so a technique known as homogenous
precipitation is employed to avoid undesirable effects. In this
method, the precipitating reagent is generated in situ
through chemical process which takes place uniformly
inside the solution. Dense precipitate is formed in this process
which can easily be filtered.
There are many different anions that can be generated in
this technique like hydroxyl (OH -) , phosphates (PO43-) ,
oxalates (C2O42-) and sulfates (SO42-). An example can be
taken as the case of precipitating hydrous iron oxide or
aluminum oxide. The precipitating reagent is produced by
the hydrolysis of urea in lower pH .
(NH2)2C=O + H2O CO2 + NH4+ + OH –
This reaction takes place gently below the boiling point of
water. Another example of homogenous precipitation is the
generation of SO42- to precipitate barium (Ba) or lead (Pb)
. The sulfate is generated in situ by heating Sulfidic acid
solution that hydrolyzes as following :
NH2SO3H + H2O H+ + SO42- + NH4+
· Experiment (1): Gravimetric determination of Cl.
Apparatus and reagents
A. 0.1N AgNO3 (dissolve 10.79 g in dilute nitric acid and
dilute to 1liter ) .
B. Conc. HNO3
C. Conc. NH3 solution
D. 3N HCl (take 252 ml of 37% HCl and dilute to liter).
E. Filtering crucibles
Procedure:
Prepare two filtering crucibles by cleaning them with
conc. HNO3 followed by washing with distilled water and
then with conc. NH3 followed by plenty of distilled water
. Record the weight of the crucibles to three decimal
places .
Take 25ml of a dilute sodium chloride solution into each
of two 250ml beaker, add excess of silver nitrate solution
slowly and constantly stirring . Heat almost to boiling for
about ten minutes. Cool the solution and add few drops of the
silver nitrate to make sure the precipitation has completed.
Let the two beakers stand still for at least one hour in a
dark place . Transfer the precipitate to the filtering crucibles
by decanting. Wash the precipitate with few ml of 6M HNO3
followed by deionized water. Continue washing until there are
no traces of AgNO3. Dry the precipitate to 110o C. for one
hour. Store the crucibles in a desiccator until they are
cooled to room temperature. Take the weight of the
crucibles and calculate the amount of chloride in the
solution. Report the % yield .
· Experiment (2): Gravimetric determination of sulfate in a
solution.
Sulfate is precipitated as barium sulfate from aqueous
solution by systematic addition of BaCl2 . Despite this
precipitation is fast and quantitative , it has great tendency
to occlude many unwanted anions such as PO43- and
NO3- .
Apparatus and reagents,
A. 0.2M BaCl2.2H2O (dissolve 48.8 g in water and dilute to
1liter) .
B. 6M HCl (half-diluted 37% HCl) .
C. 2 porcelain crucibles.
Procedure : Prepare two porcelain crucibles by cleaning and
igniting at high temperature. Cool the crucibles in a
desiccator . Weigh the crucibles repeatedly to 0.2mg. Take
two 50ml aliquots of the solution sample (SO42-) and
transfer to two 400ml beakers ; add 5ml of 6M HCl
followed by 100ml hot BaCl2 solution quickly with
vigorous stirring. Digest the precipitate for one hour. Filter the
precipitate quantitatively in an ash less filter paper. Transfer
the filter papers and the precipitates into the crucibles .Ignite
the contents until almost constant weight is obtained.
Calculate the amount of sulfate in grams.
· Experiment (3): Determination of Barium sulfate by
precipitating from homogeneous solution.
Apparatus and reagents .
A. 0.01M BaCl2.2H2O solution (A.R) [dissolve 1.22 g in
distilled water and dilute to 1liter].
B. Sulfidic acid, (NH2)2SO3H (A.R)
C. Electric Hotplate
D. Filtering crucibles (Porcelain) accurately weighed to
constant value.
Procedure : Take 100ml of the barium chloride solution
into clean dry 250ml – beaker add 1.0g of the Sulfamic
acid . Cover the beaker and heat on an electric hotplate at
980C , continue heating until turbidity begins to appear .
Filter the solution in a porcelain crucible ; wash the
precipitate with warm water twice . Heat the crucible with
precipitate at 6500C to a constant weight and determine
the weight of BaSO4 .
Titrimetric Analysis
This topic falls into a wide field that deals with
quantitative determination of an analyte , it is based on
practically reacting volumes in which the volume of a
standard reagent required to completely consume the
analyte is determined ; this procedure is known as
volumetric analysis.
Volumetric procedures employ reagents whose concentrations
are exactly known. These reagents are known as standard
solutions; the quantity of an analyte is determined from the
volume of the standard solution , this is performed by
carefully adding the standard solution into the system until
the reaction with the analyte is complete , then the volume
of the standard reagent is measured from which , the
quantitative relationship of the sought for substance is
calculated . In this procedure , the end of the analyte
reaction is determined by employing an indicating system
to denote the equivalence point during the addition of the
standard solution. This indicating technique is shown by
change in color , redox potential , or self-indicating system .
Accuracy of the volumetric analysis depends on the
accuracy of the preparation of the standard solution .
Therefore , special attention must be made to prepare
standard solution of accurate concentration . This is
achieved by dissolving a carefully weighed quantity of the
pure reagent and diluting to the desired volume . Highly
purified substance called a primary standard solution is
employed to accurately fix the real concentration of the
prepared solution . This process is known as standardization
.
Any substance that can be employed as primary standard
solution should at least has the following properties
A. It must be Commercially available in a very high purity
standard .
B. It must be least affected by heat, light, and humidity.
C. It must have high molecular weight.
Here are some of the compounds usually used as primary
standard solutions :
Substance
Molecular mass
Purity available
Standardization used
Potassium dichromate, K2Cr2O7
294.22
99.9 %
Reducing agent
Potassium hydrogen phthalate(C4H5KO4)
204.23
99.9 %
Bases
Sodium carbonate, Na2CO3
105.99
99.9 %
Acids
Potassium iodate, KIO3
214.01
99.9 %
Sodium Thiosulfate
Ethylenediaminetetraacetic Acid disodium salt, 2H2O
372.25
99 %
Metals
Sodium chloride, NaCl
58.45
99.9 %
Silver Nitrate
The importance of a titration is to find out or to estimate the
volume of a standard reagent chemically equivalent to a
given volume of an analyte solution. The point at which
the reaction stops or the analyte is theoretically consumed
is called the equivalence point. The practically observed
point in which the analyte is supposed to react completely is
called the end point. The equivalence point is a theoretical
concept while the end point is a practical concept .
In certain volumetric analysis , the end point is detected
by certain chemical substances that change in color as the
process approaches the equivalence point; these substances
are called indicators .
· Acid–base titrations.
We have studied in details the properties and reaction
of the acids and bases in chapter ( 5 ) ; but in this section
we will confine ourselves to the practical aspect of the
acid – base titrations . In acid – base titrations, given amount
of a base solution is applied to exactly determine the
chemically equivalent amount of standard acid solution , in
this reaction aqueous solution of the corresponding salt is
formed ; this process is called neutralization process.
Different indicators are used to determine the equivalence
point by changing the color of the solution . The transition
from acidic color to basic color depends on the pH . This
color change is not practically so abrupt but takes an
interval of nearly two pH units . Color change interval
varies with different indicators , so that one has to select
an indicator that distinctly changes the color at the
appropriate pH level .
In neutralization reaction :
H3+O + OH -
2H2O
This reaction denotes the neutralization of a strong acid
and strong base . From this reaction we can say that acid
is any substance that supplies hydronium ions [H+] or any
substance that consumes the hydroxide ions [OH -] ; while
the base is any substance that supplies hydroxide ions or
consumes the hydronium ions [H3O+] . This is the core of
the Arrhenius concept towards the acid – base theory.
In non-aqueous solutions , this definition is not enough
and has to modified to cope with certain reactions
involving Complexometric reactions . According to
Arrhenius , acids furnish hydronium ions in aqueous
solutions and can be stated in the following form .
HA + H2O H3+O + A - (2)
The equilibrium constant for the dissociation of the acid HA
will be:
Ka =
Assuming the concentration of H2O so large with respect
to other species , we can consider it . Therefore the
dissociation constant of the acid will be:
Ka = (3)
We also know that in aqueous solution the dissociation
constant of water ,Kw , is
Kw = [H3+O][OH -] ;
the dissociation constant of base is,
A -+ H2O HA + OH
–
Kb = (4)
From equation 3 and 4 , Ka =
Substituting for the value of Kb as , Ka = , and Kb
=
So Ka Kb = Kw , and pKa + pKb = Kw where pK
= - log k
In aqueous solutions , strong acids and strong bases
ionize completely so it becomes much easier to calculate
both hydrogen and hydroxyl ions from the stoichiometric
reaction between the acid and the base. This quantitative
process is clearly shown by using pH meter in which
stepwise addition of definite volume of the acid to the
base is recorded and the following pH curve is obtained .
Fig.1
From these curves , the equivalence point can easily be
determined . If the titration is performed without pH meter
, visual indicator is used to determine the equivalence
point .
Different acid-base titrations need different visual
indicators and choice of the indicator is determined by the
nature of the reacting acids and bases .
In strong acid-base titrations , the equivalence point fall in
between pH 3.3 and 10.7 , in that case phenolphthalein will
be the suitable indicator , see table ( 2 ) . The concentration
of the reacting species has an effect on the inflection
point. The more the concentration the sharper the inflection
point .
Experimental procedure of acid – base titrations are
conducted in this chapter. Before beginning detailed
experiment , we have to have well prepared standardized
solutions , these solutions are usually prepared from certain
substance that carry the properties of primary standard
solution . These substances include , Na2CO3 , H2C2O4 ,
H2SO4 , H3BO3 etc .
Hydrochloric acid is commonly used titrant ; although
different standard solutions can be prepared , it is more
convenient to prepare HCl of approximate normality and
standardize with a primary standard like sodium carbonate ,
Na2CO3 . The reaction takes two steps :
(a) Na2CO3 + HCl NaHCO3 + NaCl
Phenolphthalein indicator
(b) NaHCO3 + HCl CO2 + NaCl + H2O
Methyl orange indicator
Na2CO3 + 2HCl CO2 + 2 NaCl + H2O
Visual Indicators:
The main purpose of the titration is the determination of
the amount of acid or base present in any given solution .
This is achieved by locating the equivalence point which
gives us the stoichiometric amount of an acid or base
chemically equivalent to known amount of a base or an
acid .
Indicators are weak organic acids or bases that maintain a
color change in definite pH value . The pH value in which
color change takes place varies with different indicators
therefore we can select an indicator which exhibits the
color change at pH close to that obtained at the
equivalence point. We mentioned that the change from acid
color to alkaline color is not abrupt but small interval of
about two pH units involved. Considering undissociated
acid indicator as (HIn) , and undissociated base as (InOH)
and both having different colors from their ions , the
equilibria in aqueous solution will be :
HIn H + + In- and InOH
OH - + In -
By applying the law of mass action and considering the activity
coefficient as one
(constant),
Kin = and [H +] = =
this equation shows that the actual color change of the
indicator is directly related to the hydrogen-ion
concentration . Applying the definition of pH , the equation
can be written as ,
pH = - log + pKin
For weak base indicator, [OH -] = x Kn , we know
that [OH -] =
Substituting for OH -, [H+] = .
Considering acidic form of an indicator as Ina and basic form as
Inb, the equilibrium constant is expressed as :
Ina H+ + Inb , KIn = , and [H+] =
x KIn
Taking the negative logarithm , pH = pKIn + log .
Considering this above equation , the acidic color limit will
be :
pH = pKIn - log , in which pH
= pKIn – 1
the corresponding alkaline limit will be :
pH = pKIn - log , in which pH
= pKIn + 1
therefore the color change interval will be pH = pKIn
Table (2), shows different indicators with their pH range ,
color change and their preparations.
Indicator
pH - range
Acid color
Base color
Preparation
Methyl orange
3.1 – 4.4
Red
Yellow
0.01% in H2O (Na-form)
Methyl red
4.8 – 6.0
Red
Yellow
0.02g in 60ml EtOH + 40ml H2O
Phenolphthalein
8.0 – 9.6
Colorless
Red
0.05g in 50ml EtOH + 50ml H2O
Bromo cresol purple
5.2 – 6.8
Yellow
Purple
0.1g in 18.5ml of 0.01M NaOH + 225ml H2O
P-nitrophenol
5.6 – 7.6
Colorless
yellow
0.1% in H2O
Bromothymol blue
6.0 – 7.6
Yellow
Blue
0.1g in 16ml of 0.01M NaOH + 225ml H2O
Phenol red
6.4 – 8.0
Yellow
Red
0.1g in 28.2ml of 0.01M NaOH + 225ml H2O
Neutral red
6.8 – 8.0
Red
Orange
0.01g in 50ml EtOH 50ml H2O
Cresol purple
7.6 – 9.2
Yellow
Purple
0.1g in 50ml EtOH + 50ml H2O
Selected Experiments
· Acid – base titrations.
Experiment (1–1), standardization of HCl against sodium
carbonate (Na2CO3).
· Titration apparatus
A. Burette (50 ml)
B. Burette Stand
C. Pipette (10 ml)
D. Conical Flask (150 or 250 ml)
E. Methyl orange indicator solution
· Required
Solution
s.
A. 0.1N potassium hydrogen phthalate (20.3 g/liter)
B. 0.1N anhydrous sodium carbonate (Na2CO3 5.3 g/liter ) .
C. 0.1N HCl ( 8.4ml/liter )
D. 0.1N NaOH ( 4.0 g/liter) .
Procedure: Take 10 ml of the prepared sodium carbonate
into 250 ml conical flask and add 2-3 drops of methyl
orange ( yellow color will be formed ) . Use unit-volume or
graduated pipette in the process .
Fill the burette with the hydrochloric acid ; adjust the
volume reading exactly and record it . Remove any traces
of air inside the solution . Begin the titration by adding
the HCl from the burette onto the carbonate solution
carefully and systematically until the end point is close
enough . Then make the addition drop-wise until the last
drop turns the solution persistent pink color ; and record
the values . Repeat the experiment several times until the
values are close together within 2-3 units .
Take the average and report as follows:
Initial burette reading
Final burette reading
Vol. reacted, Titer (ml)
0.00
12.20
12.20
12.20
23.50
11.30
23.50
34.20
11.30
Average titer = 11.60
We know that one equivalent of any substance reacts
practically or theoretically with one equivalent of the
other. Therefore number of equivalents of Na2CO3 reacted =
Number of equivalents of HCl reacted. So that, 0.01-liter x
0.1 equiv./liter = 0.0116-liter x NHCl
NHCl = x 0.086N , which is the concentration of the
standardized hydrochloric acid .
Experiment (1-2) :
Standardization of sodium hydroxide against potassium
hydrogen phthalate(A.R), or against sulfuric acid (optional).
Reagents:
A. Titration apparatus
B. 0.1N NaOH
C. 0.1N Potassium hydrogen phthalate (C8H5KO4) or 0.1N
H2SO4
D. Phenolphthalein indicator (0.05 g in 100 ml water/ethanol
(1:1) .
Procedure: Transfer 10ml NaOH with a pipette into 250ml
conical flask . Rinse the walls of the flask with distilled
water . Fill the burette with the sulfuric acid or with the
potassium hydrogen phthalate and adjust the volume to
specific reading . Begin to add the acid into the base until
the end point gets closer ( the color is about to disappear )
. Then begin to add drop-wise until the last drop turns
the solution colorless . Tabulate the data and perform the
necessary calculations as experiment (1-1) .
Experiment (1 - 3). Determination of calcium carbonate in an
impure sample (back titration technique).
Calcium carbonate is an insoluble salt , so definite amount
of an excess acid is added to dissolve. After the formation
of a clear solution, the acid is back titrated with a base. The
sample could be limestone, marble, egg shell etc.
Requirements :
A. Titration apparatus
B. Standard 1M HCl
C. Standardized 0.1N NaOH
D. Methyl orange indicator (0.01 g – soluble M.O in water ).
E. Powdered sample
Weigh accurately 1- 1.5 g . of the carbonate sample and
transfer it to 400ml beaker. Add about 20 – 25 ml distilled
water, then add 40 ml of 1M HCl; when the effervescence
ceases completely , transfer the contents into a 100ml
volumetric flask and dilute it to the mark so that you can
establish your own dilution factors . Titrate definite
aliquots of the diluted solution with sodium hydroxide
using methyl orange indicator. calculate the amount of
calcium carbonate in the sample
CaCO3 + 2HCl CaCl2 + H2O + CO2
1 mole 2moles
M.wtCaCO3 = 100
Calculations : 100 x 100 x = % by weight of CaCO3 .
Experiment (1-4): Determination of both carbonate and
hydroxide in a mixture" commercial caustic soda analysis."
Requirements:
A. Commercial sodium hydroxide
B. Standard 0.1N HCl
C. 0.04M BaCl2 solution
D. Methyl orange and phenolphthalein indicators
In this experiment , total alkali is determined (both , CO32-
and OH-) by titrating a portion of the mixture solution
with standard acid .The other portion is added an excess
of barium chloride solution to precipitate all the carbonate
, and without filtering the solution is titrated with standard
acid ; this will give the hydroxide volume . Subtracting the
second volume from the previous titration volume , will
give the volume of the acid that neutralized the carbonate
.
BaCl2 + Na2CO3 BaCO3 + 2NaCl
V = volume of the acid using methyl orange ( total alkali )
v = volume of the acid due to phenolphthalein ( hydroxide )
then,
V – 2(V-v) = the volume of the acid that neutralized the
hydroxide. and 2(V – v) is the volume of the acid that
neutralized the carbonate .
Procedure: Prepare commercial sodium hydroxide solution by
accurately weighing about 2.5 g and transfer it quickly into
500ml volumetric flask , dissolve it well with deionized
water and fill it up to the mark . Titrate 25ml portion of
this solution with standard o.1N HCl using 3drops of methyl
orange as an indicator . Repeat the process several times
until you get almost the same reading .
Warm another 25ml portion up to 75oC and add 0.04 M
BaCl2 solution (quantitatively from burette in a slight
excess ). Cool the solution , add 2drops of phenolphthalein,
and titrate with 0.1M HCl with constant stirring until the
solution turns colorless . Repeat the process several times
to get acceptable values .
Keep in mind that 1ml of 1M HCl 0.04 g NaOH , and
1ml of 1M HCl 0.053 g Na2CO3 .
Experiment (1-5):Determination of aspirin in aspirin tablet
Aspirin is an organic compound with carboxylic and an
ester group . By hydrolyzing with an alkali , salt of weak
acid is formed which can be neutralized with a dilute
acid .
C6H4(CO2H)OOCCH3 + NaOH/H2O
C6H4(OH)CO2Na + CH3COONa
Aspirin (acetylsalicylic acid)
sodium salicylate sodium acetate
The alkali sodium acetate is titrated with dilute sulfuric
acid , using phenol red or phenolphthalein as an indicator .
Requirements:
A. Titration apparatus
B. 1M sodium hydroxide
C. 0.05M sulfuric acid
D. Phenol red or phenolphthalein indicator
E. Aspirin tablets
Exact number of aspirin tablets (1.5 – 2.0 g) are accurately
weighed and transferred into a conical flask . Add 25ml of
1M NaOH and 25ml of distilled water. Simmer the mixture
gently for about 10 – 15 minutes to hydrolyze the tablets.
After cooling the mixture , transfer quantitatively to 250ml
volumetric flask . Dilute to the mark and mix well by
repeated inversions. Titrate aliquots of the reaction mixture
with standard 0.05M H2SO4 using phenol red or
phenolphthalein .
· Calculate the weight of acetylsalicylic acid .
· Molecular wt. of acetylsalicylic acid = 180.16.
· 1mol of H2SO4 1mol of acetylsalicylic acid.
Experiment (1-6):Determination of available nitrogen in
fertilizers.
If ammonium salts are treated with an excess of strong
alkali like sodium hydroxide ammonia gas is liberated. The
gas is then trapped in excess of dilute acid which can be
back titrated with dilute sodium hydroxide.
NH4Cl + NaOH heat NH3 + NaCl + H2O , or
NH4+ + OH- NH3 + H2O
Keep in mind that nitrogen compounds are essential nutrients in
most fertilizers.
Requirements:
A. Distillation assembly
B. Titration apparatus
C. Standard 0.1N sodium hydroxide
D. Standard 0.1N hydrochloric acid
E. Methyl orange indicator
F. Fertilizer sample (or NH4Cl salt)
Procedure: The distillation apparatus is set up in such a
way that the receiver adapter is immersed below the
surface of the hydrochloric acid .Weigh accurately definite
amount of fertilizer (or NH4Cl salt) , not more than 2.0 g ,
into 50ml distillation flask . Add 25ml of 2N NaOH
solution , few boiling chips and quickly connect to the
distillation apparatus. Heat the contents gently to get
smooth boiling. Avoid any sucking back from the receiver
adapter during boiling . Continue the distillation process
until about 10ml of the original solution remains ( 30 – 40
min) .
When the distillation is over , disconnect the apparatus
from top before removing the Bunsen burner or any other
heating system . Wash any residues in the condenser and
receiver adapter into a 150ml conical flask and titrate
against .0.1N NaOH using 3drops of methyl orange
indicator.
Calculate:
A. The weight of ammonia distilled.
B. Percent of available nitrogen in the fertilizer.
· Precipitation Titrations
Precipitation titrations are among the oldest analytical methods
employed in chemical analysis. Up to now they are applied in
certain volumetric analysis such as the determination of
silver , chlorides , bromides , iodides and thiocyanates. In this
method , certain organic indicators are used ; these
indicators have the property of adsorption or desorption on
the solid materials formed during the precipitation . This
adsorption and desorption process takes places near the
equivalence point accompanied by change in the color ; the
procedure is called Fagan’s method as an honour for Polish
scientist Kazimierz Fagan’s ; among these indicators are
fluorescein and its derivatives . Since most of these
procedures involve the application of silver nitrate
solutions they are called argentometric methods.
We have mentioned that the most important precipitation
titration utilizes silver nitrate as the reagent for the
process ; therefore in these titrations , we will theoretically
confine our discussions to argentometric reactions .Before
we launch the practical aspect of this titration , let us
consider certain important concepts relating the solubility
of the salt AgCl which forms during the titration process.
Silver chloride is sparingly soluble in water, and considering
its (AgCl) saturated solution the solubility equilibrium will
be represented as:
AgCl(s) Ag (aq) +
Cl -(aq)
Ksp =
This is an example of heterogeneous equilibrium so the
concentration of the solid part (AgCl) remains constant in
the solution and can be taken as unity (1) so that :
Ksp = [Ag+][Cl-]
This molar product of Ag+ and Cl - at equilibrium is
called solubility product . In expressing the solubility
product of the ions involved , the stoichiometric
coefficients are raised .
Ag2S 2Ag+ + S2- ,
Ksp = [Ag+]2[S2-]
Table 2.2 , shows the solubility product (Ksp) of certain
slightly soluble salts at 25oC .
It must be understood that the smaller the value , the less
soluble salt in water . We can determine the Ksp from the
molar concentration at equilibrium and vice versa . Ksp is
comparatively easy to calculate with respect to other
equilibrium calculations .
Example 1.
An aqueous saturated solution of silver carbonate was
analysed and found to contain 8 mg of the salt dissolved
in 250 ml. Calculate the Ksp of the Ag2CO3 .

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  • 1. Chemistry of life (Biochemistry) The study of chemical compounds that are vital for living organisms to sustain life is called biochemistry. The subject deals with the nature of these compounds and characteristic reactions they make inside the living organisms . We are not involved fundamentally with the study of biochemistry as a subject , but to give brief introduction to main classes of the organic compounds in this important field. It is beyond this discussion to present detailed explanation of these essential organic substances . We will give short introduction of the main classes and their active role in our body . Some of these groups are , carbohydrates , fats and proteins, etc.. · Carbohydrates. Carbohydrates are classes of organic compounds that consist of carbon , hydrogen and oxygen with an empirical formula of Cm(H2O)n in most cases . The terms m and n can be the same as in the case of C6H12O6 (glucose) or different in the case of C12H22O11 (sucrose) . Another important feature of the carbohydrates is that oxygen and hydrogen are generally in ratio of 2:1 , so that it was historically called hydrates of carbon ; but not all compounds of carbohydrates necessarily maintain this hydrogen – oxygen ratio and not all compounds that fit this hydrogen-oxygen ratio are carbohydrates . In biochemistry the term carbohydrate denotes different compounds called saccharides . These compounds include sugars , starch and cellulose . Saccharides (Greek word meaning sugars) are generally classified into monosaccharides , disaccharides and polysaccharides . Monosaccharides are the simple sugars which are either aldoses (aldehydes) like glucose or ketoses ( ketones) like
  • 2. fructose . These simple sugars are further classified on the base of the number of carbon atoms they contain like pentose (containing five carbon atoms) , or hexose (containing six carbon atoms) . Carbohydrates are naturally formed in a process called photosynthesis in which plants combine CO2 from the air and water from the soil in the presence of chlorophyll , sunlight and certain enzymes producing simple sugars . 6 CO2 + 6H2O (sun light) C6H12O6 + 6O2 sugar(glucose) This above reaction is not simple process as it looks , but extremely complicated reaction with different intermediate steps before it gives the final product . since the final product is a monosaccharide , plants have the ability to synthesize disaccharides by combining two molecules of monosaccharides . 2 C6H12O6 C12H12O11 + H2O monosaccharide disaccharide This above natural chemical process is the reverse of hydrolysis . Both plants and animals have the ability to synthesize polysaccharides by combining large number of monosaccharide molecules . n C6H12O6 (C6H10O5)n + n H2O polysaccharide in plants are usually in the form of cellulose , stored in stalks and stems and in a form of starch in the roots and seeds . Animals and plants depend on each other to sustain life ; plants can synthesize carbohydrates - as we mentioned
  • 3. before - from simple inorganic materials like CO2 and H2O while animals cannot. On the other hand , animals utilize these essential carbohydrates and metabolically oxidize to give CO2 , water and energy . C6H12O6 + 6 O2 6 CO2 + 6H2O + energy Note that this reaction is the reverse of that of the photosynthesis in plants ; we can combine both equations to establish the relationship . The reaction to the right is endothermic and the energy is taken from the sunlight while the reaction to the left is exothermic in which the same energy is liberated and utilized by the body activity to sustain life . It is clear from this cycle that the energy stored in the carbohydrates is originally from the sun .This energy is used by all living organisms in their metabolic process. 1- Monosaccharides Monosaccharides are the basic units of the carbohydrates ; they are also called simple sugars . The term monosaccharide originally came from two Greek words mono- which means single and Sachar which means sugar . These simple sugars are usually colorless crystalline solid material . Almost all monosaccharides are soluble in water and have sweet taste .
  • 4. Most monosaccharides have chemical formula of Cm(H2O)n where n < 2 . They are classified - as we already described - on the base of number of carbon atoms present in the molecule such as triose , pentose , and hexose representing glyceraldehyde , ribose and glucose respectively . The carbon chain of the monosaccharides are numbered beginning from the carbon closest to the carbonyl group . We can also designate the structural nature of the monosaccharide by using the terms aldoses or ketoses so that if the carbonyl is at position 1 of the carbon chain , the sugar is an aldehyde and termed an aldose . On the other hand if the carbonyl group is in between two carbon atoms , then the sugar is ketone and termed ketose . · Structures of monosaccharides – (optical isomers). Monosaccharides exist in different stereo-isomeric forms . In section (10.3) of this chapter , we presented this property in detail and explained the two types of the stereoisomers ; the important one in our study of simple sugars is the optical isomerism . Optical isomers are molecules that have the same molecular formula but differ in the arrangement of the atoms in space in such a way that they are mirror images to one another . Despite these isomers have the same physical and chemical properties , they differ in the direction of rotating planely polarized light (light waves vibrating in one plane ) . If polarized light is passed through these isomers, the plane of the vibrating light is
  • 5. rotated due to presence of chiral or asymmetric carbon in the molecule that is a carbon atom attached to four different groups . Suitable example is the glyceraldehyde molecule . The central carbon atom is connected to aldehyde group , OH group , hydrogen and alcoholic group . Despite glyceraldehyde is the simplest carbohydrate , it is very important compound in the since that many complex sugars can be derived . · Classification of monosaccharides We have seen that simple sugars can be divided into different groups depending on the number of carbon atoms they contain designating them as trioses , tetroses, pentoses and hexoses. A. Trioses: These are monosaccharides that consist of three carbon atoms . Glyceraldehyde is an example of this group . Trioses are the product of the metabolic breakdown of certain simple sugars . B. Tetroses: Tetrodes are simple sugars with four-carbon structure . Erythrosine and throes are examples of these sugars .
  • 6. C. Pentose: These are monosaccharides with five-carbon atoms . They are classified into two groups ; those with aldehyde functional groups called aldopentose and those with ketone functional groups called ketopentose . The most significant of these sugars are the ribose and deoxyribose which are found in RNA (Ribonucleic acid) and in DNA (Deoxyribonucleic acid) . Both RNA and DNA are the main component of every living cell , especially in the nucleus of the cell . D. Hexoses: They are monosaccharides with six carbon atoms . There are different kinds of hexoses but the most important ones in terms of human body are glucose , fructose and galactose . These sugars have the same chemical formula but differ in structure and can be called isomers of a given molecular formula . We already explained that simple sugars are classified on the base of their functional groups . Hexose with aldehyde functional group are called aldohexose and those with ketone functional group are called ketohexose . A. Glucose: glucose is an aldohexose with general formula C6H12O6 ; there are four asymmetric carbon atoms and 16
  • 7. optical isomers . Glucose molecules can exist in solution in the form of open chain or in the form of ring structure . This ring structure is formed when the hydrogen of the carbon 5 -OH attaches with the oxygen atom of the aldehyde group on carbon 1 . Open chain Ring structure B. Galactose: galactose is an aldose sugar like glucose ; it can be considered as the isomer of glucose , they only differ in the configuration of a single carbon atom which is known as epimoric form of isomerism (differ in one stereoisomeric center) . Like any hexose , it can exist in solution as an open chain or ring form . Open chain Ring structure C. Fructose: fructose is ketohexose with a molecular formula of C6H12O6 , like glucose and galactose . Fructose or levulose is naturally occurring sugar found in fruits ; like any other hexoses , it is found in solution as a ring
  • 8. or as open chain structure . Fructose is the sweetest and most soluble of all sugars . Reactions of the hexose monosaccharides Aldoses and ketoses have reducing properties in common . These properties are the bases of the chemical tests undertaken to determine the sugar content in human blood and urine . The reactions involved in these chemical test are in the organic practical section . In these tests aldoses are oxidized into their corresponding acids . RCHO + Cu2+ + 2 OH- RCOOH + Cu2O + H2O Clinitest tablets are used to perform the above test to determine the sugar in the human urine . Note that these tablets contain copper ions in the form of Cu(II)sulfate. Another test for reducing sugars is the Tollens test in a solution containing silver ions in basic solution . Glucose + Ag+ + OH- gluconic acid + Ag + water
  • 9. Another general qualitative test for carbohydrates is the Molisch test in which concentrated sulfuric acid is carefully added to the testing solution containing alpha- naphthol ; formation of purple ring at the interface of the acid and solution confirms the presence of carbohydrate substance . Another important reduction reaction for some aldoses like glucose is the fermentation process in which an enzyme from the yeast catalyzes this reaction and without which this reaction cannot take place . C6H12O6 enzyme CH3CH2OH + CO2 Aldoses can also undergo oxidation since they both contain aldehyde and hydroxyl group . We have seen that oxidation of the aldehyde group gives the corresponding acid , like gluconic acid in the case of glucose and glucuronic acid if the hydroxyl group oxidizes . Aldohexoses can be converted to alcohols by reduction and in this case , glucose is reduced to sorbitol and fructose is reduced to mixture of mannitol and sorbitol. Glucose Sorbitol Since both sorbitol and mannitol are partially and slowly
  • 10. taken by the body , it is useful as sweetening agent for the diabetic people . When orally taken some of the sugar is excreted in the urine and other is taken by the body slowly . 1. Disaccharides Disaccharides are the sugars formed when two monosaccharides are chemically joined together . The most common examples of the disaccharides are sucrose , lactose and maltose . They all have the general formula C12H22O11 , which means they are isomers . Disaccharides formed as a result of condensation reaction of two monosaccharides which means elimination of water molecule . The reverse reaction is known as hydrolysis performed by certain enzyme . These reactions are vital in the metabolism of living things . Different disaccharides hydrolyze to give two different or same simple sugars as shown above. Disaccharides are , like monosaccharides , white crystalline sweet solids . Their solubility varies from completely soluble to slightly soluble . Disaccharide molecule from two simple sugars is formed by displacing hydroxyl group from one molecule and the hydrogen ion from the other producing water molecule a process known as condensation reaction . Sucrose is formed by the reaction of the aldehyde group
  • 11. of glucose with the ketone group of fructose thus eliminating the reducing property of the carbonyl; and for this reason give negative test to reducing reagents. On the other hand , maltose and lactose have free aldehyde group and give positive test for both Fehling and Tollens reagents . In the case of maltose formation , one aldehyde group of a glucose reacts with the hydroxyl group of another glucose leaving one aldehyde group free for reducing property . In the same way , lactose has one available aldehyde group ; for that reason maltose and lactose are reducing sugars while sucrose is not reducing sugar . These sugars are differentiated analytically on the base of their fermentation and reducing properties . Both sucrose and maltose can be fermented by the addition of yeast which contains sucrase and maltase enzymes . Lactose does not ferment with the yeast since it does not contain lactase . A. Sucrose: This sugar is known as table sugar ; it is naturally occurring carbohydrate found in many fruits and vegetables like carrot roots and pineapples . Sucrose is extracted from cane and beet sugars and refined to be consumed by the people. Hydrolysis of sucrose gives a mixture of glucose and fructose . This mixture of equal amounts of glucose and fructose is invert sugar . High concentration of this invert sugar is found in honey ; this kind of sugar is less prone to crystallization when cooled .
  • 12. B. Maltose: Maltose is a disaccharide formed from two units of glucose . It is commonly known as malt sugar . It is commercially prepared from hydrolysis of starch . It generally occurs in germinating seeds . C. Lactose: Lactose is also a disaccharide formed from joining of two units of glucose and galactose . It is found in milk and that is the reason why it is called milk sugar ; it differs from the above sugars in the sense that it comes from animals. We pointed out previously that lactose is fermented by the enzyme lactase forming lactic acid; this fermented milk is said to be sour milk. Lactose is also found in urine of pregnant woman and since it is reducing sugar , it gives positive test with Fehling or Benedict solution. 2. Polysaccharides: Polysaccharides can be defined as the polymeric form of monosaccharides . These polymers hydrolyze to yield many monosaccharide molecules . These polysaccharides are generally formed from five-carbon or six-carbon sugars . The most common polysaccharides are : A. Starch Starch is a polymer of glucose sugars which consist of around 20% amylose and 80% amylopectin . Amylose is linear chain of glucose molecules while amylopectin is branched form of the starch and contain much larger units of glucose in comparison to amylose . Starches are insoluble in water .
  • 13. Starch (long chain of glucose units ) Amylopectin (branched form of starch) Amylose (linear chain of glucose) The presence of starch is tested with iodine which gives characteristic deep blue color . Hydrolysis of starch to simple sugars through several stages giving different dextrin’s on the process which can be followed by addition of iodine . Iodine gives no color with simple sugars and blue color with dextrin’s . A. Cellulose Cellulose is also polysaccharide consisting of linear chain of several hundred to thousands of linked glucose units . It is an important supporting and structural component of all plants . Unlike starch , cellulose is not affected by the digestive human enzymes and for that reason helps prevent constipation by making bulk feces of dietary fiber when taken by human .
  • 14. Cellulose Cellulose is digested and metabolized by certain animals like ruminants and termites with the help of micro- organisms which live in their abdomen . Cellulose is not soluble in water and in majority of solvents . Unlike starch , it does not give positive test to iodine or to Fehling reagent . Materials like cotton , wood and paper are mainly composed of cellulose . Cellulose is used to produce paper , cellophane and rayon. B. Glycogen , Glycogen is multi-branched glucose polymer which is responsible for the storage of energy in animals . Glycogen is produced by the body and stored in the liver and the muscles ; it is originated from the animals while starch is basically from plant origin ; it acts as a reserve or long- term energy storage . Even though fats act as the primary energy stores , glycogen energy is necessary for immediate mobilization to cover a sudden need for glucose. Glycogen is synthesized in the body cell from glucose through a process called glycogenesis and hydrolyzed to give glucose through a process known as glycogenolysis . Glucose glycogenesis
  • 15. Glycogen glycogenolysis Glucose C. Dextrin Dextrin’s are lower molecular weight carbohydrates formed as a result of starch hydrolysis . Dextrin’s are considered as an intermediate between the starch and the disaccharides . It dissolves in water giving sticky colloidal suspension used in the preparation of adhesives like the glue on the back of the postage stamp . 1. Lipids (fats) Lipids are naturally occurring organic compounds ; these include fats , waxes , fat soluble vitamins and sterols (steroid alcohols) . It is one of the main sources of food essential for animals and plants . Lipids consist of carbon , hydrogen and oxygen and in certain cases nitrogen and phosphorus . They are insoluble in water but soluble in organic solvents like alcohol and acetone . Even though there are different categories of lipids , we will confine our discussion to simple lipids which are esters of fatty acids . · Fatty acids: There are two types of fatty acids ; saturated fatty acids which contain single bond between the carbon atoms and unsaturated fatty acids which contain double between the carbon atoms .
  • 16. CH3-CH2-CH2-CH2-COOH CH3-(CH2)7- CH=CH-(CH2)7-COOH Saturated fatty acid unsaturated fatty acids Table.10.6 Common saturated fatty acids. Name Formula Source Butyric acid C3H7COOH Butter fat Caproic acid C5H11COOH Goat fat Caprylic acid C7H15COOH Coconut oil Capric acid C9H19COOH Palm oil Lauric acid C11H23COOH Laurel (plant) Table.10.7 Common unsaturated fatty acids . Name Formula Source Oleic acid C17H33COOH (1 double bond) Olive oil Linoleic oil C17H31COOH (2 double bond) Linseed oil
  • 17. Linolenic acid C17H29COOH (3 double bond) Linseed oil Unsaturated fatty acids can exist in two structural forms , cis in which the two hydrogen atoms adjacent to the double bond are on the same side of the carbon chain and trans in which the adjacent two hydrogen atoms are on the opposite sides of the carbon atom (discussed in section 10.3) . Iodine number Human body cannot sufficiently synthesize some of the fatty acids they need and must be obtained from the food they take . These fatty acids are known as essential fatty acids . linoleic and linolenic acids are among the essential acids ; they are found in certain plant oils like peanut , soybeans and corn but not in olive oil or cocoanut. Essential fatty acids have important function in the body such as the synthesis of prostaglandins (hormone-like substance with wide physiological activities) . Iodine number: The degree or the amount of unsaturation in a given fatty acid is determined by its iodine number , which is the amount of iodine consumed by 100 g of the acid. Iodine reacts with the double bond of the unsaturated fatty acid , so the higher the iodine consumed the more double bonds present in that fatty acid .
  • 18. Despite there are different methods for determining iodine value , they all fall under the iodiometry procedures . We indicated that the higher the unsaturation for a fatty acid the higher the iodine value . Linseed oil has much more iodine value than coconut oil which means the linseed oil is highly unsaturated oil with respect to coconut oil . In general , vegetables oils have higher iodine number than animal fats . Fats and oils We defined the fats as esters of fatty acids (already discussed in carboxylic acids unit) which are formed from the reactions between a fatty acid and specific alcohol called glycerol . There are three OH groups available in glycerol and each one can react with a fatty acid molecule forming a fat molecule . These fatty acids can be of same fatty acids or different fatty acid molecules .
  • 19. The esters formed when the alcohol is glycerol is called glyceride . It can be monoglyceride , diglyceride or triglyceride depending on the number of fatty acids reacted with the glycerol . In general if the reacting fatty acid is saturated , the ester formed is called fat and if it is unsaturated the resulting ester will be an oil . Most fats have iodine value less than 70 while most oils have iodine value more than 70 . Keep in mind that mineral oil is a saturated hydrocarbon different than fats and oils which are either from animal , vegetable origin and petroleum distillate . Those oils are different from essential oils which are volatile liquids used as flavors and perfumes , while mineral oils are mixtures of higher alkanes . The name may be inappropriate and sometimes other names are applied such as liquid paraffin , paraffin oil etc. . · Properties and uses of fats Fats can be considered as body fuel , they produce more energy for the body than any other nutrient such as carbohydrates or proteins . Another important function of the fats is their essential role in reserving food supply for the body . They protect the vital organs by keeping them rigid in their place and helping to absorb shocks . In terms of physical properties , fats and oils are either in liquid or solid state with white or yellow color in most cases . They are colorless and tasteless in their pure state but get rancid as the time goes on creating unpleasant odor and taste. Since both fats and oils are insoluble in water ; they form transient emulsions which can be stabilized by adding emulsifiers from the bile to make them digestible . In terms of chemical properties , fats hydrolyze in the
  • 20. presence of certain enzymes to form fatty acids and glycerol . We have seen that when a fat is formed water is the product (see the preceded equation) ; the reverse process is called hydrolysis. After the hydrolysis of the fat , the glycerol is separated and purified for medical and industrial purposes . It is used as humectant (against loss of moisture) for skin and hair care products . It is also used in the preparation of medical and pharmaceutical products . Glycerol has also an important industrial applications ; it is found in food , beverages and sweetener as sugar substitute besides many other uses . We briefly explained in this chapter , that basic hydrolysis of fats yield glycerol and salt of the fatty acid ; this process is called saponification and the sodium or potassium salt formed is called soap . Sodium soaps are solid bars and potassium soaps are liquid . In preparing soaps, various materials are added to give pleasant odor and color ; in certain soaps , germicidal substances are added to protect the skin from the germs as in the case scouring soap . In hard water , calcium and magnesium ion react with the soap rendering it insoluble and inactive to give more
  • 21. lather and more cleansing result . This problem is overcome by using detergents . Detergents are synthetic cleaning substances consisting of mixtures of surfactants (lower the surface tension) used as a soap . They can be used both in soft and hard water ; which means calcium and magnesium salts of detergents are soluble in water thus overcoming the disadvantage of soap . Detergents are made from different petrochemicals besides fat and oils . They are generally neutral unlike the soaps which are alkaline . The sodium salt of the detergents is prepared by the reaction of long chain alcohols with concentrated sulfuric acid and the product is neutralized by sodium hydroxide . R-OH + H2SO4 R- OSO3H + H2O long chain alcohol R- OSO3H + NaOH R-OSO3Na + H2O Detergent The unbranched straight chain detergents are less harmful in terms of environmental pollution . 2. Proteins Another essential nutrient for living organisms is the protein . Proteins are complex organic substance which consist of carbon , hydrogen , oxygen and nitrogen besides some other elements like sulfur and phosphorus . Proteins consist of long chain of simple molecules called amino acids arranged in a specific order. There are 20 known amino acids . All of these amino acids cannot be synthesized by the human body and must be externally supplied . The first
  • 22. three amino acids are shown below with their symbol and names Amino acid molecule contains amino group and carboxylic group . Despite the amino group can be structurally anywhere in the molecule , the amino group of the naturally occurring amino acids are bonded with the alpha – carbon (the one next to acid group ). Almost all amino acids have alpha chiral carbon (except glycine) and show L-configuration . Amphoteric nature of the amino acids Amphoteric substance is one that shows both acidic and basic character ; which means it behaves as an acid and base. As the name denotes , amino acids contain carboxylic group which is an acid since it furnishes proton in solution and also contain amino group which accepts the proton and on the base of Brnsted – Lowry acid-base definition , amino acids act as an acid and as a base at the same time; therefore amino acids react with both acidic and basic substances ; consequently amino acids can be called amphoteric substance as long as they donate and accept protons .
  • 23. Amino acid molecules react together in which the amino group of one amino acid reacts with the carboxylic group of another forming a long chain of amino acid molecules connected through a bond known as peptide bond . This bond is formed as a result of condensation reaction in which one molecule of water is produced for every two reacting amino acids . A combination between two amino acids form a product called dipeptide , and that of three amino acids form tripeptide and more than three amino acids form polypeptide. There are two different ways for two amino acids to combine . It is very important to know that for combined amino acids , their name is abbreviated by taking the first three letters of the amino acid and in the order of their combination in the peptide ; the first amino acid is the
  • 24. one that furnishes the OH from the acid group . · Isoelectric point We have seen that amino acids contain both basic and acidic functional groups. In the crystalline forms , amino acids exist as zwitterions , which is a neutral molecule having both positive and negative electrical charges resulting no net electric charge. These solid forms usually have high melting point and less solubility in organic solvents but soluble in water . In aqueous solution , these amino acids establish an equilibrium dependent on the pH of the solution . isoelectric point In basic solution , the equilibrium is shifted towards the right and in acidic solution the equilibrium is shifted towards the left . If two electrodes are placed in this amino acid solution , there will be net migration of the solute molecules towards either the positive electrode (cathode) or the negative electrode (anode) depending on the alkalinity or acidity of the solution . In basic solution a negatively charged carboxylic group is formed and the migration takes place towards the positively charged electrode and in acidic solution , a positively charged amino group will be formed and the migration takes place towards the negatively charged electrode .
  • 25. At certain pH , characteristic of each amino acid , the positive and the negative ions will be equal and there will be no net migration of the amino acid molecules towards the electrodes . This specific pH value is called isoelectric point . Proteins like amino acids have an isoelectric point which is characteristic property for each protein . Proteins have the minimum solubility in their isoelectric point . For the pH of the amino acid above the isoelectric point , the basicity dominates , that is there will be more basic sites and the pH below the isoelectric point , the acidity will dominate and there will be more acidic sites . · Structure and the properties Proteins hydrolyze in acidic or basic media , or by the action of certain enzymes . This hydrolysis breaks the protein molecule into the constituent amino acids . In the reverse process , amino acids combine together with the help of specific enzymes to form polypeptides which are the basic molecular structures of the proteins ; the more units of amino acids combined the bigger the molecular weight of the protein . As a result of the large molecular sizes of the proteins , different structural forms are created which are referred to as primary , secondary and tertiary structures based on the shape and spatial orientation of the protein molecule . The bases of these shapes and their biological significances is beyond our discussion. The main properties of the proteins can be summarized as follows :
  • 26. A. Colloidal The aqueous solution of protein form colloidal dispersion of the protein molecules in water . These colloidal particles have the ability to pass through the filter paper but inable to pass thhrough the cellular membranes ; this phenomenon is physiologically important since it keeps the protein in the bloodstream and impedes the diffusion of protein molecules into the urine indicating the damage of kidney membranes . B. Denaturation Denaturation refers to change of protein structure loosing its biological function . When proteins become denaturated , they coagulate or precipitate . This effect takes place under the influence of several reasons : 1. Effect of alcohol: 70% aqueous solution of alcohol (ethanol) precipitates almost all proteins and for this reason we use this alcohol concentration as a disinfectant because it coagulates the protein of the bacteria rendering it inactive . 2. Salting out process: at high concentration of salt solution , proteins become almost insoluble and precipitate . This is one of the techniques used to separate proteins from solutions . Salts like Na2SO4 , NaCl are used for this purpose . The separation from the solution is achieved through filteration and the traces of the remaining salt is eliminated by a process known as dialysis . 3. Heavy metal salt : Saturated salt solutions of certain heavy metals such as silver and mercury(II) precipitate the protein . These heavy metal salts coagulate the protein
  • 27. making it biologically inactive . This is the reason why these metals are poisonous if taken internally . 4. Heating : Almost all protein coagulate on heating . Heating is common method to sterilize materials because the protein of the bacteria is destroyed by coagulating it . Presence of protein in urine (which is kidney malfunction) can be determined by heating given sample of urine to precipitate the protein . 5. Concentrated inorganic acids: Concentrated strong mineral acids can be used to precipitate the protein. HCl , H2SO4 , and nitric acid are among the mineral acids that can be employed to precipitate proteins . Concentrated HCl from the gastric glands precipitate the milk protein , casein , in the form of curds . C. Qualitative tests for proteins Most qualitative tests for proteins depend on specific color formation . These tests are not always decisive since it depends on the presence of certain amino acids . Some of these tests are : Xanthoprotic : Which is the formation of yellow precpitate due to action of nitric acid on proteins . Contact of nitric acid on your finger usually produces yellow color on the skin of the finger . This test is only positive for the amino acids that contain benzene ring . There are other well known tests like , Biuret , Millon’s , Hopkin-Cole and ninhydrin tests in which their procedures are explained in the organic practical section. Different
  • 28. chromatographic methods are used to separate and identify different mixture of proteins or amino acids . Quantatively the amount of protein in a sample is estimated by determining the nitrogen content and multiplying the result by factor 6. This factor came from the fact that the average percent of nitrogen in protein is 16% by weight ; that is approximately 1/6 of the total weight of the protein . If for example the total nitrogen of a sample is 2.5 g , the amount of protein present in the sample = . The result is always expressed in percent . Nucleoproteins Nucleoproteins can be defined as conjugated propteins that consist of acidic part which is nonprptein section called nucleicacid and a protein part . Nucleoproteins hydrolyze during the digestion process into nucleic acids and proteins . Proteins further hydrolyze into amino acids while nucleic acids hydrolyze into nucleotides . Nucleic acids are biological polymers that consist of repeating units of nucleotides so that nucleic acids can be called polynucleotides . Each nucleotide consist of heterocyclic nitrogen base , pentose sugar and phosphoric acid . The number of nucleotide units in a nucleolic acid depends on the nature of the nucleic acid and may range
  • 29. from approximately one hundred units to several million units . Nucleotide There are mainly two branches of nucleic acids : a) Deoxyribonucleic acid (DNA) which is found in the nucleus of the cell and contains pentose sugar , deoxyribose , which is the only sugar on hydrolysis. b) Ribonucleic acid (RNA) which is mainly found in cytoplasm and contain ribose as the only sugar on hydrlysis . Parts of nucleotides We have showed that a nucleotide consist of nitrogen base , pentose sugar (ribose or deoxyribose ) and phosphoric acid . Let us investigate them one by one . A. Nitrogen bases: The two groups of heterocyclic nitrogen – containing bases in the nucleic acids are derivatives of purine or pyrimidine with following structures .
  • 30. The most common purine bases in nucleic acids are adenine and quinine . They are present in both DNA and RNA . The pyrimidines that are most common in both DNA and RNA are cytosine , uracil and thymine . (2-oxy-6-aminopyrimidine) (2,4-dioxy-5- methylpyrimdine) (2,dioxypyrimidine) Nucleosides , Nucleosides are units formed by combination of nitrogenous base and one of the two pentose sugars (ribose or deoxyribose) in the structure of nucleic acids . In this combination the glycosidic bond between the sugar and the nitrogenous base is formed at C – 1 of the sugar and the position 9 of the purine base or position 1 of the pyrimidine base .
  • 31. Glyosidic between C1 of ribose Glyosidic between C1 of deoxyribose and and positon-9 of purine postion-1 of pyrimidine Nomenclature Nucleosides from the pyrimidines have names ending in - idine and those from the purines have names ending in - osine Purines with ribose sugars are named as follows : Adenine + ribose Adenosine Guanine + ribose Guanosine Purines with deoxyribose sugars are named as follows : Adenine + deoxyribose Deoxyadenosine Guanine + deoxyribose Deoxy guanosine Pyrimidines containing ribose sugars are named as follows : Uracil + ribose Uridine Cytosine + ribose Cytidine Thymine + ribose Thymidine Pyrimidines containing deoxyribose sugars are named as : Uracil + deoxyribose Deoxy uridine Cytosine + deoxyribose Deoxycytidine Thymine + deoxyribose Deoxythymidine
  • 32. It is known that glycosides are quite stable in alkaline medium and the situation is the same in the nucleosides but hydrolyze in aqueous acid solution to produce base and pentose sugar . Polynucleotides Hydrolyze Nucleotides Hydrolyze Nucleosides Hydrolyze Base + sugar Nucleotides are the phosphate esters of the nucleosides . The phosphoric acid reacts with one of the hydroxyl groups in the sugar forming phosphate ester bond. Condensation reaction In the case of ribonucleoside , the phosphate ester bond can be formed at three possible -OH groups at C1 , C3 and C5 and for that reason we have to specify the point of the attachment in naming the nucleotide . We already pointed out that nucleotides are phosphate esters of nucleosides ; and because the compound contains phosphoric acid , acidic name is given according to the following table. Nucleoside + H3PO4 Acidic name of the nucleotide Adenosine Adeynilic acid Guanosine Guanilic acid Cytidine Cytidilic acid
  • 33. Uridine Uridylic acid Thymidine Thymidilic acidic The nucleoside formed from the reaction of one molecule of a nucleoside with one molecule of a phosphoric acid is called monophosphate of that nucleoside . In the case of adenosine , it is called adenosine monophosphate (AMP) or Adeynilic acid . If it reacts with one more phosphoric acid molecule , it is called adenosine diphosphate (ADP) ; and if three groups are involved it is called adenosine triphosphate (ATP) . ) AMP) (ADP) (ATP) These phosphate esters , AMP , ADP and ATP perform vital role in cellular metabolism as high storage of energy and release that energy from the hydrolysis of phosphate bonds and pass to the specific reactions that require this energy to take place . ATP contains the highest amount of energy in comparison to ADP and AMP. Besides being body building blocks , certain nucleotides perform biological activities of their own . Appropriate example can be taken in the case of cyclic adenosine monophosphate (cyclic – AMP) which is formed as a result of phosphate group attached to carbon-3 and carbon-5 of the ribose sugar . It (cyclic-AMP) acts as messenger in regulating enzymatic activities in cells that store fats and carbohydrates.
  • 34. Cyclic adenosine monophosphate (camp) Certain nucleotides act as Coenzyme . Nicotinamide adenine dinucleotide (NAD+) is essential for many biological oxidation-reductions reactions to take place . It exists in oxidized form (NAD+) and reduced form transferring electrons from reaction to another . Exercises: 1. What is the modern definition of organic chemistry ? Briefly explain the role of organic chemistry in our daily life. 2. What was the vital force theory and how was it disproved ? 3. Account for the fact that element carbon has the unique property of forming so many millions of chemical compounds . 4. Briefly compare , in general , the physical and chemical properties of organic and inorganic compounds . 5. Explain the role of carbon dioxide in the natural formation of certain organic substances like coal , petroleum , and natural gas . 6. What are the hydrocarbons ? mention the different classes of the hydrocarbons on the base of their general structure . Give an example of each class . 7. Name the following hydrocarbon compounds .
  • 35. 8. Write the structural formula for each of the following compounds . A. 3-Methylheptane B. 2,3-Dimethylpentane C. 2-Chloro-3-ethylpentane D. 1,1,2,2-Tetrabromobutane E. e) 2-Butyne F. 3,4-Diethyl-1-hexyne G. 3-Ethyl-2-heptene 9. Write the possible isomers for the following hydrocarbons . A. C7H16 B. C6H12 C. C5H10 D. C4H8 E. C6H10 10. How can you determine the nature of the following hydrocarbons as alkanes, alkenes, alkynes and cycloalkanes . A. C7H14 B. C6H12 C. C5H12 D. C4H6 E. C3H4 11. How can you distinguish by simple qualitative test between the following pairs of compounds ? A. Ethanol and pentane .
  • 36. B. Acetaldehyde and acetone . C. Acetic acid and ethylamine . D. Phenol and benzene . E. Pentane and pentene . 12. Account for the extra stability of benzene in comparison to ethene (ethylene). 13. Explain what we mean by a functional group . Why it is important to classify organic compounds on the base of their functional group ? 14. Distinguish the following compounds as alcohol , aldehyde , ketone , carboxylic acid and amine . A. CH3-COCH3 B. CH3CH(OH)CH3 C. HCHO D. CH3CH2CO2H E. CH3CH(NH2)CH3 . 15. Draw the possible structural formula of the following compounds . A. C2H6O B. CH4O C. C3H6O2 D. C3H8O E. C2H7N. 16. Complete and balance the following equations .
  • 37. 17. Define the following terms . A. Fatty acids B. Esters C. Oils D. Acid chlorides 18. Give an example of each of the following reactions . A. Addition reaction B. Substitution reaction C. Elimination reaction 19. What are nucleoproteins ? 20. Explain briefly the hydrolysis product of nucleic acid . 21. What is the basic difference between the RNA and DNA in terms of structure ? 22. Write T at the end of the true statement and F at the end of the false one. A. Adenosine consist of ribose sugar and pyrimidine base . B. Hydrolysis of nucleotide gives phosphoric acid and nucleoside . C. The nucleoside urosine is made up of uracil and ribose . D. The nitrogen containing bases in the nucleic acids are either derivatives of pyrimidine or pyridine . E. Uracil is found in both RNA and DNA .
  • 38. 23. Explain the relationship between the plants and animals in terms of synthesis and utilization of carbohydrates . 24. Despite that lactose and sucrose have the same formula (C12H22O11), they give different results to reducing agents . Explain why ? 25. Write (√ ) at the end of the correct statement and (X) at the of the incorrect one . A. Carbohydrates were historically known as hydrates of carbon . ( ) B. Carbohydrates are synthesized by plants and animals from inorganic materials . ( ) C. Proteins are essential for living organisms as source of energy only . D. Alkaline hydrolysis of fats gives fatty acids . E. Proteins are polymeric units of amino acids . F. Large value of iodine number for fats shows more unsaturated sites of that fat . G. All the compounds in which the ratio of C to H is 2:1 are necessarily carbohydrates . H. Nucleosides do not contain phosphate units. 26. Explain the following terms : a) Fatty acids b) Isoelectric point c) Saturated fatty acids d) Detergents e) peptide bond.
  • 39. 27. Give reasons for the following statements. A. All the naturally occurring amino acids are optically active except glycine . B. 70% aqueous ethanol is better disinfectant than 100% ethanol (absolute) . C. Chiral compounds are optically classified as D and L forms in their aqueous solutions . D. Pentose and hexose sugar molecules can exist as ring form structure . E. Enantiomers are different than epimers . C C CH 2 OH OH OHH C C CH 2 OH OH HOH D -glyceraldehydeL-glyceraldehyde CHO C C CH 2
  • 44. + 3H 2 O glycerol fatty acids triglyceride Fatty acid+alcohol Fat+H 2 O esterification Hydrolysis H 2 N - C - H COOH H H 2 N - C - H CH 3 COOH H 2 N - C - H CH 2 OH COOH Glycine Alanine Serine H 2
  • 45. N - C - COOH R H Amino group Carboxylic group HOOC - C - NH 2 H H +HCl HOOC - C - NH 3 + +Cl - H H NH 2 - C- COOH + NaOH NH 2 - C- COO- + Na + + H 2 O H H H H reactas base reactas an acid H 2 N - CH - C
  • 46. R 1 O NH - CH - COOH R 2 Peptide bond H 2 N - CH 2 - C O OH + H 2 N - CH - C O OH NH 2 - CH 2 - C -NH - CH - COOH O CH 3 Glycine Gly-ala + H 2 O CH 3 Alanine O
  • 47. OH H 2 N - CH 2 - C O OH Glycine NH 2 - CH - C - NH - CH 2 - COOH CH 3 O H 2 N - CH - CH3 + Alanine Ala-gly H 2 O + CH 3 6 CO 2 + 6H 2 O + ENERGY plant photosynthesis. animal metabolism
  • 48. C 6 H 12 O 6 +6O 2 CH 3 CH 3 Br CH 3 - CH 2 - CH - CH - CH 3 A- B- C- CH 3 COOH + CH 3 CH 2 OH C 3 H 8 + O 2 CH
  • 50. OH OHH H C C C C C CH 2 OH OH OHH HOH H OHH OH O D-(+)-Glucose D-(-)-Fructose D-(+)-Galactose Practical Section A. Wet chemistry practical’s (inorganic portion) In the following practical work , we will be confined to experiments based on wet chemistry or bench chemical analysis ; these are traditional laboratory tests which encompasses certain processes like sampling , weighing , preparing solutions and performing classical chemical tests . In ordinary wet chemistry analysis , qualitative and quantitative results can be obtained , yet it is rarely employed in areas like industry , research centers , forensic analysis etc. These institutions employ modern chemical analysis which is extension of the classical analytical techniques that has been automated and computerized to
  • 51. save time and increase the sensitivity selectivity of the system . The following laboratory sessions involves certain quantitative topics like gravimetry , titrimetry and qualitative topics on identification methods based on formation of color , precipitate or evolution of specific gas . Practical chemistry is mere laboratory work in which the theoretical knowledge of the subject is developed and the necessary skills are acquired . To undertake this work , the necessary apparatus , equipment and reagents must be available and become familiar with the students . For any given experiment , these materials must be planned and prepared so that the result can be reported in a scientific manner . The students must have knowledge of the application of these tools and get acquainted with the laboratory environment . We can classify the laboratory tools into two parts: A. Glassware: which are glass containers used for mixing , measuring , holding and heating laboratory reagents ; they are generally made from durable and chemically inert materials like borosilicate’s . These glass containers are of different shapes and sizes used for different purposes. Some of these laboratory glassware are stated below . 1. Beakers : Beakers are glass containers of almost cylindrical shape used for heating mixing and holding samples . They are available in different sizes . Volumetric Flaska Elenmeyer Flaska
  • 52. 2. Flasks : Flasks are narrow-necked and wider bottom glass containers used for measuring preparing and heating solutions . Volumetric flasks are used for preparing and diluting solutions of definite concentrations . Erlenmeyer flasks or conical flasks are used for heating or handling solutions . Boiling Flasks 3. Boiling flasks: flat bottom flasks used for heavy duty boiling. They are available in different forms and shapes according to their application. Test tubes Different boiling flasks Test tubes holders 4. Test tubes: Small glass tubes resistant to high temperature used for testing samples especially qualitative tests .
  • 53. Beakers 5. Funnels : Funnels are laboratory apparatus with a wide mouth and long pipe like opening at the end used to transfer liquid substance or finely grinded particles to avoid spillage . They are usually made of glass , plastic or ceramic . Different funnels 6. Burettes: Burettes are long graduated glass tubes with stopcock at end for drawing accurate amount of a solution . It is extensively used in analytical chemistry, there are different sizes and structures Different burettes 7. Other laboratory glassware: Pipettes are used for taking fixed volume of a liquid. They are usually made of glass or plastic materials.
  • 54. Uni-Volume Pipette Graduated Pipettes Pipette fillers Crucibles wash bottles Desiccator Glass rods Burette clamp Burette stand Test tube rack B. Instruments: The other feature of the laboratory equipment are the instruments. Instruments are devices that help or facilitate to undertake experimental work . meter hotplate electronic balance Mortar and pestle Bunsen burners Oven Shaker
  • 55. Gravimetric analysis One of the most accurate and precise methods in chemical analysis is the gravimetric analysis . In this method , the analyte is converted to insoluble form by precipitating from the solution . The precipitate is then separated , washed , dried or ignited , weighed and calculated . Perfect example can be taken in the determination of Cl- content in a given solution by precipitating as AgCl with silver nitrate solution . After filtering and drying , the precipitate is weighed and the chloride content is calculated . There are different forms of gravimetric analysis , and the one we are concerned in this discussion is the one in which the sought for substance is almost completely precipitated from a solution so that no considerable amount is lost during filtration , washing and weighting .To achieve successful analytical result , the following points has to be fulfilled : A. The precipitated substance must be sufficiently insoluble so that we can analytically ignore the dissolved amount . B. The precipitate particles must be large enough to be quantitatively filtered and the impurities can easily be washed . C. The final precipitate must be a substance of definite chemical composition after physical or chemical treatments. · To accomplish these points and obtain almost fine precipitate particles, the following problems must be overcome too. 1. Co-precipitation, the precipitate formed may contain foreign impurities from the solution which depends on the nature of the precipitate and the conditions in which the precipitate was formed ; this contamination can be due to either the adsorption of impurities on the surface of the precipitated particles in the solution or the occlusion of foreign
  • 56. particles during the process of crystallization . 2. Coagulation, is the tendency of adsorption layers of the precipitate to attract ions of opposite charge forming large size particles containing solvent molecules . 3. Occlusion, this is the way in which foreign particles are trapped inside the precipitate during the crystallization . It is not easy to get rid of the occluded impurities . 4. Post-precipitation, it is the precipitation that takes place on the surface of the precipitating reagent , usually it comes from a substance that has common ion with primary precipitate , e. g CaC2O4 / Mg++ & C2O4- - . Treatment of the Precipitate To prepare relatively fine pure precipitate , the following favorable conditions has to be fulfilled. A. The precipitation must be implemented in dilute solution keeping in mind the solubility of the precipitate and digestion period , to minimize the co-precipitation. B. Add the reagents together slowly with constant stirring to create large crystals. C. Precipitation is done in hot solution provided no counter effect in terms of solubility and the stability of the precipitate is resulted. D. The precipitate should be washed with a suitable solvent. Precipitation from Homogenous solution We have already seen that to create a desirable precipitate , we must minimize the degree of supersaturation so that dilute solution of the precipitating agent is gently and slowly added to the sample solution with a constantly effective stirring . Even though these techniques maintain a low degree of supersaturation yet excess of the precipitating reagent is unavoidable so a technique known as homogenous precipitation is employed to avoid undesirable effects. In this method, the precipitating reagent is generated in situ through chemical process which takes place uniformly
  • 57. inside the solution. Dense precipitate is formed in this process which can easily be filtered. There are many different anions that can be generated in this technique like hydroxyl (OH -) , phosphates (PO43-) , oxalates (C2O42-) and sulfates (SO42-). An example can be taken as the case of precipitating hydrous iron oxide or aluminum oxide. The precipitating reagent is produced by the hydrolysis of urea in lower pH . (NH2)2C=O + H2O CO2 + NH4+ + OH – This reaction takes place gently below the boiling point of water. Another example of homogenous precipitation is the generation of SO42- to precipitate barium (Ba) or lead (Pb) . The sulfate is generated in situ by heating Sulfidic acid solution that hydrolyzes as following : NH2SO3H + H2O H+ + SO42- + NH4+ · Experiment (1): Gravimetric determination of Cl. Apparatus and reagents A. 0.1N AgNO3 (dissolve 10.79 g in dilute nitric acid and dilute to 1liter ) . B. Conc. HNO3 C. Conc. NH3 solution D. 3N HCl (take 252 ml of 37% HCl and dilute to liter). E. Filtering crucibles Procedure: Prepare two filtering crucibles by cleaning them with conc. HNO3 followed by washing with distilled water and then with conc. NH3 followed by plenty of distilled water . Record the weight of the crucibles to three decimal places . Take 25ml of a dilute sodium chloride solution into each of two 250ml beaker, add excess of silver nitrate solution slowly and constantly stirring . Heat almost to boiling for about ten minutes. Cool the solution and add few drops of the
  • 58. silver nitrate to make sure the precipitation has completed. Let the two beakers stand still for at least one hour in a dark place . Transfer the precipitate to the filtering crucibles by decanting. Wash the precipitate with few ml of 6M HNO3 followed by deionized water. Continue washing until there are no traces of AgNO3. Dry the precipitate to 110o C. for one hour. Store the crucibles in a desiccator until they are cooled to room temperature. Take the weight of the crucibles and calculate the amount of chloride in the solution. Report the % yield . · Experiment (2): Gravimetric determination of sulfate in a solution. Sulfate is precipitated as barium sulfate from aqueous solution by systematic addition of BaCl2 . Despite this precipitation is fast and quantitative , it has great tendency to occlude many unwanted anions such as PO43- and NO3- . Apparatus and reagents, A. 0.2M BaCl2.2H2O (dissolve 48.8 g in water and dilute to 1liter) . B. 6M HCl (half-diluted 37% HCl) . C. 2 porcelain crucibles. Procedure : Prepare two porcelain crucibles by cleaning and igniting at high temperature. Cool the crucibles in a desiccator . Weigh the crucibles repeatedly to 0.2mg. Take two 50ml aliquots of the solution sample (SO42-) and transfer to two 400ml beakers ; add 5ml of 6M HCl followed by 100ml hot BaCl2 solution quickly with vigorous stirring. Digest the precipitate for one hour. Filter the precipitate quantitatively in an ash less filter paper. Transfer the filter papers and the precipitates into the crucibles .Ignite the contents until almost constant weight is obtained.
  • 59. Calculate the amount of sulfate in grams. · Experiment (3): Determination of Barium sulfate by precipitating from homogeneous solution. Apparatus and reagents . A. 0.01M BaCl2.2H2O solution (A.R) [dissolve 1.22 g in distilled water and dilute to 1liter]. B. Sulfidic acid, (NH2)2SO3H (A.R) C. Electric Hotplate D. Filtering crucibles (Porcelain) accurately weighed to constant value. Procedure : Take 100ml of the barium chloride solution into clean dry 250ml – beaker add 1.0g of the Sulfamic acid . Cover the beaker and heat on an electric hotplate at 980C , continue heating until turbidity begins to appear . Filter the solution in a porcelain crucible ; wash the precipitate with warm water twice . Heat the crucible with precipitate at 6500C to a constant weight and determine the weight of BaSO4 . Titrimetric Analysis This topic falls into a wide field that deals with quantitative determination of an analyte , it is based on practically reacting volumes in which the volume of a standard reagent required to completely consume the analyte is determined ; this procedure is known as volumetric analysis. Volumetric procedures employ reagents whose concentrations are exactly known. These reagents are known as standard solutions; the quantity of an analyte is determined from the volume of the standard solution , this is performed by carefully adding the standard solution into the system until the reaction with the analyte is complete , then the volume of the standard reagent is measured from which , the quantitative relationship of the sought for substance is
  • 60. calculated . In this procedure , the end of the analyte reaction is determined by employing an indicating system to denote the equivalence point during the addition of the standard solution. This indicating technique is shown by change in color , redox potential , or self-indicating system . Accuracy of the volumetric analysis depends on the accuracy of the preparation of the standard solution . Therefore , special attention must be made to prepare standard solution of accurate concentration . This is achieved by dissolving a carefully weighed quantity of the pure reagent and diluting to the desired volume . Highly purified substance called a primary standard solution is employed to accurately fix the real concentration of the prepared solution . This process is known as standardization . Any substance that can be employed as primary standard solution should at least has the following properties A. It must be Commercially available in a very high purity standard . B. It must be least affected by heat, light, and humidity. C. It must have high molecular weight. Here are some of the compounds usually used as primary standard solutions : Substance Molecular mass Purity available Standardization used Potassium dichromate, K2Cr2O7 294.22 99.9 % Reducing agent Potassium hydrogen phthalate(C4H5KO4) 204.23 99.9 %
  • 61. Bases Sodium carbonate, Na2CO3 105.99 99.9 % Acids Potassium iodate, KIO3 214.01 99.9 % Sodium Thiosulfate Ethylenediaminetetraacetic Acid disodium salt, 2H2O 372.25 99 % Metals Sodium chloride, NaCl 58.45 99.9 % Silver Nitrate The importance of a titration is to find out or to estimate the volume of a standard reagent chemically equivalent to a given volume of an analyte solution. The point at which the reaction stops or the analyte is theoretically consumed is called the equivalence point. The practically observed point in which the analyte is supposed to react completely is called the end point. The equivalence point is a theoretical concept while the end point is a practical concept . In certain volumetric analysis , the end point is detected by certain chemical substances that change in color as the process approaches the equivalence point; these substances are called indicators . · Acid–base titrations. We have studied in details the properties and reaction of the acids and bases in chapter ( 5 ) ; but in this section we will confine ourselves to the practical aspect of the acid – base titrations . In acid – base titrations, given amount
  • 62. of a base solution is applied to exactly determine the chemically equivalent amount of standard acid solution , in this reaction aqueous solution of the corresponding salt is formed ; this process is called neutralization process. Different indicators are used to determine the equivalence point by changing the color of the solution . The transition from acidic color to basic color depends on the pH . This color change is not practically so abrupt but takes an interval of nearly two pH units . Color change interval varies with different indicators , so that one has to select an indicator that distinctly changes the color at the appropriate pH level . In neutralization reaction : H3+O + OH - 2H2O This reaction denotes the neutralization of a strong acid and strong base . From this reaction we can say that acid is any substance that supplies hydronium ions [H+] or any substance that consumes the hydroxide ions [OH -] ; while the base is any substance that supplies hydroxide ions or consumes the hydronium ions [H3O+] . This is the core of the Arrhenius concept towards the acid – base theory. In non-aqueous solutions , this definition is not enough and has to modified to cope with certain reactions involving Complexometric reactions . According to Arrhenius , acids furnish hydronium ions in aqueous solutions and can be stated in the following form . HA + H2O H3+O + A - (2) The equilibrium constant for the dissociation of the acid HA will be: Ka = Assuming the concentration of H2O so large with respect
  • 63. to other species , we can consider it . Therefore the dissociation constant of the acid will be: Ka = (3) We also know that in aqueous solution the dissociation constant of water ,Kw , is Kw = [H3+O][OH -] ; the dissociation constant of base is, A -+ H2O HA + OH – Kb = (4) From equation 3 and 4 , Ka = Substituting for the value of Kb as , Ka = , and Kb = So Ka Kb = Kw , and pKa + pKb = Kw where pK = - log k In aqueous solutions , strong acids and strong bases ionize completely so it becomes much easier to calculate both hydrogen and hydroxyl ions from the stoichiometric reaction between the acid and the base. This quantitative process is clearly shown by using pH meter in which stepwise addition of definite volume of the acid to the base is recorded and the following pH curve is obtained . Fig.1 From these curves , the equivalence point can easily be determined . If the titration is performed without pH meter , visual indicator is used to determine the equivalence point . Different acid-base titrations need different visual indicators and choice of the indicator is determined by the nature of the reacting acids and bases .
  • 64. In strong acid-base titrations , the equivalence point fall in between pH 3.3 and 10.7 , in that case phenolphthalein will be the suitable indicator , see table ( 2 ) . The concentration of the reacting species has an effect on the inflection point. The more the concentration the sharper the inflection point . Experimental procedure of acid – base titrations are conducted in this chapter. Before beginning detailed experiment , we have to have well prepared standardized solutions , these solutions are usually prepared from certain substance that carry the properties of primary standard solution . These substances include , Na2CO3 , H2C2O4 , H2SO4 , H3BO3 etc . Hydrochloric acid is commonly used titrant ; although different standard solutions can be prepared , it is more convenient to prepare HCl of approximate normality and standardize with a primary standard like sodium carbonate , Na2CO3 . The reaction takes two steps : (a) Na2CO3 + HCl NaHCO3 + NaCl Phenolphthalein indicator (b) NaHCO3 + HCl CO2 + NaCl + H2O Methyl orange indicator Na2CO3 + 2HCl CO2 + 2 NaCl + H2O Visual Indicators: The main purpose of the titration is the determination of the amount of acid or base present in any given solution . This is achieved by locating the equivalence point which gives us the stoichiometric amount of an acid or base chemically equivalent to known amount of a base or an acid . Indicators are weak organic acids or bases that maintain a color change in definite pH value . The pH value in which color change takes place varies with different indicators
  • 65. therefore we can select an indicator which exhibits the color change at pH close to that obtained at the equivalence point. We mentioned that the change from acid color to alkaline color is not abrupt but small interval of about two pH units involved. Considering undissociated acid indicator as (HIn) , and undissociated base as (InOH) and both having different colors from their ions , the equilibria in aqueous solution will be : HIn H + + In- and InOH OH - + In - By applying the law of mass action and considering the activity coefficient as one (constant), Kin = and [H +] = = this equation shows that the actual color change of the indicator is directly related to the hydrogen-ion concentration . Applying the definition of pH , the equation can be written as , pH = - log + pKin For weak base indicator, [OH -] = x Kn , we know that [OH -] = Substituting for OH -, [H+] = . Considering acidic form of an indicator as Ina and basic form as Inb, the equilibrium constant is expressed as : Ina H+ + Inb , KIn = , and [H+] = x KIn Taking the negative logarithm , pH = pKIn + log . Considering this above equation , the acidic color limit will be : pH = pKIn - log , in which pH = pKIn – 1 the corresponding alkaline limit will be : pH = pKIn - log , in which pH = pKIn + 1
  • 66. therefore the color change interval will be pH = pKIn Table (2), shows different indicators with their pH range , color change and their preparations. Indicator pH - range Acid color Base color Preparation Methyl orange 3.1 – 4.4 Red Yellow 0.01% in H2O (Na-form) Methyl red 4.8 – 6.0 Red Yellow 0.02g in 60ml EtOH + 40ml H2O Phenolphthalein 8.0 – 9.6 Colorless Red 0.05g in 50ml EtOH + 50ml H2O Bromo cresol purple 5.2 – 6.8 Yellow Purple 0.1g in 18.5ml of 0.01M NaOH + 225ml H2O P-nitrophenol 5.6 – 7.6 Colorless
  • 67. yellow 0.1% in H2O Bromothymol blue 6.0 – 7.6 Yellow Blue 0.1g in 16ml of 0.01M NaOH + 225ml H2O Phenol red 6.4 – 8.0 Yellow Red 0.1g in 28.2ml of 0.01M NaOH + 225ml H2O Neutral red 6.8 – 8.0 Red Orange 0.01g in 50ml EtOH 50ml H2O Cresol purple 7.6 – 9.2 Yellow Purple 0.1g in 50ml EtOH + 50ml H2O Selected Experiments · Acid – base titrations. Experiment (1–1), standardization of HCl against sodium carbonate (Na2CO3). · Titration apparatus A. Burette (50 ml) B. Burette Stand C. Pipette (10 ml) D. Conical Flask (150 or 250 ml) E. Methyl orange indicator solution · Required
  • 68. Solution s. A. 0.1N potassium hydrogen phthalate (20.3 g/liter) B. 0.1N anhydrous sodium carbonate (Na2CO3 5.3 g/liter ) . C. 0.1N HCl ( 8.4ml/liter ) D. 0.1N NaOH ( 4.0 g/liter) . Procedure: Take 10 ml of the prepared sodium carbonate into 250 ml conical flask and add 2-3 drops of methyl orange ( yellow color will be formed ) . Use unit-volume or graduated pipette in the process . Fill the burette with the hydrochloric acid ; adjust the volume reading exactly and record it . Remove any traces of air inside the solution . Begin the titration by adding the HCl from the burette onto the carbonate solution carefully and systematically until the end point is close enough . Then make the addition drop-wise until the last drop turns the solution persistent pink color ; and record the values . Repeat the experiment several times until the values are close together within 2-3 units .
  • 69. Take the average and report as follows: Initial burette reading Final burette reading Vol. reacted, Titer (ml) 0.00 12.20 12.20 12.20 23.50 11.30 23.50 34.20 11.30 Average titer = 11.60 We know that one equivalent of any substance reacts practically or theoretically with one equivalent of the other. Therefore number of equivalents of Na2CO3 reacted = Number of equivalents of HCl reacted. So that, 0.01-liter x 0.1 equiv./liter = 0.0116-liter x NHCl NHCl = x 0.086N , which is the concentration of the standardized hydrochloric acid . Experiment (1-2) : Standardization of sodium hydroxide against potassium
  • 70. hydrogen phthalate(A.R), or against sulfuric acid (optional). Reagents: A. Titration apparatus B. 0.1N NaOH C. 0.1N Potassium hydrogen phthalate (C8H5KO4) or 0.1N H2SO4 D. Phenolphthalein indicator (0.05 g in 100 ml water/ethanol (1:1) . Procedure: Transfer 10ml NaOH with a pipette into 250ml conical flask . Rinse the walls of the flask with distilled water . Fill the burette with the sulfuric acid or with the potassium hydrogen phthalate and adjust the volume to specific reading . Begin to add the acid into the base until the end point gets closer ( the color is about to disappear ) . Then begin to add drop-wise until the last drop turns the solution colorless . Tabulate the data and perform the necessary calculations as experiment (1-1) . Experiment (1 - 3). Determination of calcium carbonate in an impure sample (back titration technique). Calcium carbonate is an insoluble salt , so definite amount
  • 71. of an excess acid is added to dissolve. After the formation of a clear solution, the acid is back titrated with a base. The sample could be limestone, marble, egg shell etc. Requirements : A. Titration apparatus B. Standard 1M HCl C. Standardized 0.1N NaOH D. Methyl orange indicator (0.01 g – soluble M.O in water ). E. Powdered sample Weigh accurately 1- 1.5 g . of the carbonate sample and transfer it to 400ml beaker. Add about 20 – 25 ml distilled water, then add 40 ml of 1M HCl; when the effervescence ceases completely , transfer the contents into a 100ml volumetric flask and dilute it to the mark so that you can establish your own dilution factors . Titrate definite aliquots of the diluted solution with sodium hydroxide using methyl orange indicator. calculate the amount of calcium carbonate in the sample CaCO3 + 2HCl CaCl2 + H2O + CO2 1 mole 2moles M.wtCaCO3 = 100 Calculations : 100 x 100 x = % by weight of CaCO3 .
  • 72. Experiment (1-4): Determination of both carbonate and hydroxide in a mixture" commercial caustic soda analysis." Requirements: A. Commercial sodium hydroxide B. Standard 0.1N HCl C. 0.04M BaCl2 solution D. Methyl orange and phenolphthalein indicators In this experiment , total alkali is determined (both , CO32- and OH-) by titrating a portion of the mixture solution with standard acid .The other portion is added an excess of barium chloride solution to precipitate all the carbonate , and without filtering the solution is titrated with standard acid ; this will give the hydroxide volume . Subtracting the second volume from the previous titration volume , will give the volume of the acid that neutralized the carbonate . BaCl2 + Na2CO3 BaCO3 + 2NaCl V = volume of the acid using methyl orange ( total alkali ) v = volume of the acid due to phenolphthalein ( hydroxide ) then, V – 2(V-v) = the volume of the acid that neutralized the hydroxide. and 2(V – v) is the volume of the acid that neutralized the carbonate .
  • 73. Procedure: Prepare commercial sodium hydroxide solution by accurately weighing about 2.5 g and transfer it quickly into 500ml volumetric flask , dissolve it well with deionized water and fill it up to the mark . Titrate 25ml portion of this solution with standard o.1N HCl using 3drops of methyl orange as an indicator . Repeat the process several times until you get almost the same reading . Warm another 25ml portion up to 75oC and add 0.04 M BaCl2 solution (quantitatively from burette in a slight excess ). Cool the solution , add 2drops of phenolphthalein, and titrate with 0.1M HCl with constant stirring until the solution turns colorless . Repeat the process several times to get acceptable values . Keep in mind that 1ml of 1M HCl 0.04 g NaOH , and 1ml of 1M HCl 0.053 g Na2CO3 . Experiment (1-5):Determination of aspirin in aspirin tablet Aspirin is an organic compound with carboxylic and an ester group . By hydrolyzing with an alkali , salt of weak acid is formed which can be neutralized with a dilute acid .
  • 74. C6H4(CO2H)OOCCH3 + NaOH/H2O C6H4(OH)CO2Na + CH3COONa Aspirin (acetylsalicylic acid) sodium salicylate sodium acetate The alkali sodium acetate is titrated with dilute sulfuric acid , using phenol red or phenolphthalein as an indicator . Requirements: A. Titration apparatus B. 1M sodium hydroxide C. 0.05M sulfuric acid D. Phenol red or phenolphthalein indicator E. Aspirin tablets Exact number of aspirin tablets (1.5 – 2.0 g) are accurately weighed and transferred into a conical flask . Add 25ml of 1M NaOH and 25ml of distilled water. Simmer the mixture gently for about 10 – 15 minutes to hydrolyze the tablets. After cooling the mixture , transfer quantitatively to 250ml volumetric flask . Dilute to the mark and mix well by repeated inversions. Titrate aliquots of the reaction mixture with standard 0.05M H2SO4 using phenol red or phenolphthalein . · Calculate the weight of acetylsalicylic acid . · Molecular wt. of acetylsalicylic acid = 180.16.
  • 75. · 1mol of H2SO4 1mol of acetylsalicylic acid. Experiment (1-6):Determination of available nitrogen in fertilizers. If ammonium salts are treated with an excess of strong alkali like sodium hydroxide ammonia gas is liberated. The gas is then trapped in excess of dilute acid which can be back titrated with dilute sodium hydroxide. NH4Cl + NaOH heat NH3 + NaCl + H2O , or NH4+ + OH- NH3 + H2O Keep in mind that nitrogen compounds are essential nutrients in most fertilizers. Requirements: A. Distillation assembly B. Titration apparatus C. Standard 0.1N sodium hydroxide D. Standard 0.1N hydrochloric acid E. Methyl orange indicator F. Fertilizer sample (or NH4Cl salt) Procedure: The distillation apparatus is set up in such a way that the receiver adapter is immersed below the
  • 76. surface of the hydrochloric acid .Weigh accurately definite amount of fertilizer (or NH4Cl salt) , not more than 2.0 g , into 50ml distillation flask . Add 25ml of 2N NaOH solution , few boiling chips and quickly connect to the distillation apparatus. Heat the contents gently to get smooth boiling. Avoid any sucking back from the receiver adapter during boiling . Continue the distillation process until about 10ml of the original solution remains ( 30 – 40 min) . When the distillation is over , disconnect the apparatus from top before removing the Bunsen burner or any other heating system . Wash any residues in the condenser and receiver adapter into a 150ml conical flask and titrate against .0.1N NaOH using 3drops of methyl orange indicator. Calculate: A. The weight of ammonia distilled. B. Percent of available nitrogen in the fertilizer. · Precipitation Titrations Precipitation titrations are among the oldest analytical methods employed in chemical analysis. Up to now they are applied in certain volumetric analysis such as the determination of silver , chlorides , bromides , iodides and thiocyanates. In this method , certain organic indicators are used ; these
  • 77. indicators have the property of adsorption or desorption on the solid materials formed during the precipitation . This adsorption and desorption process takes places near the equivalence point accompanied by change in the color ; the procedure is called Fagan’s method as an honour for Polish scientist Kazimierz Fagan’s ; among these indicators are fluorescein and its derivatives . Since most of these procedures involve the application of silver nitrate solutions they are called argentometric methods. We have mentioned that the most important precipitation titration utilizes silver nitrate as the reagent for the process ; therefore in these titrations , we will theoretically confine our discussions to argentometric reactions .Before we launch the practical aspect of this titration , let us consider certain important concepts relating the solubility of the salt AgCl which forms during the titration process. Silver chloride is sparingly soluble in water, and considering its (AgCl) saturated solution the solubility equilibrium will be represented as: AgCl(s) Ag (aq) + Cl -(aq) Ksp = This is an example of heterogeneous equilibrium so the
  • 78. concentration of the solid part (AgCl) remains constant in the solution and can be taken as unity (1) so that : Ksp = [Ag+][Cl-] This molar product of Ag+ and Cl - at equilibrium is called solubility product . In expressing the solubility product of the ions involved , the stoichiometric coefficients are raised . Ag2S 2Ag+ + S2- , Ksp = [Ag+]2[S2-] Table 2.2 , shows the solubility product (Ksp) of certain slightly soluble salts at 25oC . It must be understood that the smaller the value , the less
  • 79. soluble salt in water . We can determine the Ksp from the molar concentration at equilibrium and vice versa . Ksp is comparatively easy to calculate with respect to other equilibrium calculations . Example 1. An aqueous saturated solution of silver carbonate was analysed and found to contain 8 mg of the salt dissolved in 250 ml. Calculate the Ksp of the Ag2CO3 .