UNITI Laboratory Safety

Accidents
lean and experiment.
Pharmaceutical Chemistry laboratory is a place to
carelessness but can be prevented by strictly
following the instructions
are usually
caused by from injury in
the general safety rules to help and guide students
given by staff. Following are

a laboratory.
activity without teacher's
1. Laboratory is a serious place to work. Do not perform any
permission.
be worn when working in the Laboratory.
2. Laboratory apron must safety masks should
such as safety goggles, gloves,
3. Personal Protective Equipments (PPE)
be used whenever necessary.

4. Long hair should be tied

back to reduce possibility ofcatching fire. safe in the
feet at all times. Bare feet or
use ofsandals are not
5. Wear shoes that cover the
laboratory.
bottles before removing the
contents.
6. Check the labels of Pour a small amount ofchemical
droppers into the chemical bottles.
7. Donot insert pipettes or

into the glass container for

use.
bulb for
Never draw chemicals into
a pipette with the mouth/use
8. Never taste any chemical.
pipetting. in the laboratory.
and smoking is prohibited
9. Eating, drinking, chewing gums area immediately
with
contact with eyes or skin, wash the
into
10.If any chemical
comes

sufficient quantity ofwater. flames.

chemicals away from open
flammable and combustible
11.Keep
combustible gases only
under the directions ofyour teacher. Use the
12.Handle toxic and
materials are present.
fume hood when such done only
unused chemicals and products ofreaction should be
13.Disposal of broken glass,
as directed by the teacher. tube should not be pointed at
in a test tube, the mouth oftest
14.When heating a substance
another person or yourself.
shower and first aid kit.
15.Know the location of fire extinguisher, safety
the laboratory.
water before leaving
16.Wash hands with soap and concerned
smoke is observed, immediately inform the
17.Ifanyunusual noise or smell, or

staff
/1
Pharmaceutical Organic Chemistry-I
A Practical Book of
Unit-I
 Experiments based on
UNITII Laboratory Techniques
A) Recrystallization
relies on the fact that solubility increases as
Principle: Recrystallization (or crystallization)
saturated solution c0ols, it becomes supersaturated and the
temperature increases. As hot,
a
an impure (crude) solid
out. In a recrystallization procedure,
solute precipitates (crystallizes)
solution is cooled, one pure product from the crude solid
is dissolved in a hot solvent. As this
crystallizesout while the other stays dissolved.

Procedure:
Benzoic acid in 30-35 ml ofhot
Using ahotplate, dissolve approximately 1.0g of impure
water (water at or near its boiling point.)
in a 125 ml Erlenmeyer flask. Ifthere is a residual
a small amount of additional solvent (H,O)
amount ofmaterial that does not dissolve, adding
as it is important to use minimum amount
of
is essential. Do not continue to add more solvent,
insoluble in the hot solvent and will
solvent during recrystallization. This material is probably
Ifthere is any such insoluble solid
be separated from the hot solution by gravity filtration.
Preheat the funnel by running small
residue, filter this hot solution through a fluted filter paper.
acid in the
amount of hot water in it. This may reduce the risk of crystallization of Benzoic
stem ofthe funnel, or on filter paper, and so reduce its loss. Allow the filtrate to cool gradually
to room temperature. When the cooled solution drops to, or near, room temperature, coolit
further in an ice-water bath. Collect the resulting crystals by vacuum filtration.
Transfer the

crystals from the filter paper to a tared watch glass and let it dry thoroughly before weighing
Determine the amount of Benzoic acid recovered and, based uponthe initial amount with
which the experiment started, determine the percentage ofrecovery.
Observations:
Initial amount ofimpure Benzoic acid(g)=W
i)Tare weight ofwatch glass(g)= W,
i)Weight of watch glass + purecrystals(g)=W,
iv) Amount of Benzoic acid recovered W,-W,w, =

General Guidelines:
1)Choose appropriate solvent(s) such that,
product is very soluble in it at high temperatures

2/Unit-II
 low temperatures
product is not soluble in it at
impurities are either soluble at all temperatures
or

insoluble at all temperatures (can be filtered off

solid
2) Dissolve impure for reference)
mass (also take a melting point
Weigh out crude solid and record its
Add a boiling chip or boiling stick (otherwise, may
it "bump"and spill)
add more)
solvent (add a bit, heat/swirl. Ifnot dissolved,
Use a minimum amount of hot
no crystals.
NOTE: Too much solvent will produce
INeeded: Decolorize
3) OnlyIf
colorless solutions (looks like water)
Most pure compounds are white give
and
which adsorbs
To remove color, which are reallytrace
contaminants, add activated charcoal,
the impuriies.

4) Only If Needed:
Use fluted filter paper and a hot, stemless funnel
will crystallize out and be lost.
NOTE: fthe solution cools, product
rinse flask, then filter
Use a small amount ofhot solvent to
RECRYSTALLIZATION part
5) Crystallize solute to get the 0° C (put in ice-water bath)
Cool the solution slowly: hot (boiling) room temperature
NOTE: This gives large, pure crystals
and leaves impurities in solution.

A Practical Book of Pharmaceutical Organic Chemistry-II /3

B) Steam Distillation
Steam distillation is a means of separating and purifying organic compounds. This involves
volatilizing a substance by passing steam into a mixture of the compound and water. Many
organic compounds tend to decompose at high- sustained temperatures. This is used when
separation by normal distillation would not be possible. In this procedure water or steam is
introduced into the distillation apparatus. By adding water or steam, the boiling points of
compounds are depressed, allowing them to evaporate at lower temperatures, preferably
below the temperatures at which the deterioration ofthematerial becomes appreciable.After
distillation the vapours are condensed as usual, usually giving a two- phase system of water
and the organic compounds, allowing for simple separation.

Applications
For the separation of the desired organic compound:
(a) from non-volatile tarry substances, which are formed as by-products in many reactions,
(b) from aqueous mixtures containing dissolved inorganic salts;
(c)in cases where otherprocedures ofseparation might pose difficulties (e.g. the direct ether
extraction of aniline, produced by the reduction of nitrobenzene by tin, etc., leads to troublesome
emulsion formation owing to the alkali and the tin compounds present)
(d)fromcompounds that are not appreciably volatile in steam (eg.o-nitrophenol from
p-nitrophenol) and
(e) from certain by-products that are steam volatile (e.g. biphenyl and excess of unreacted
starting materials from the less volatile triphenylcarbinol).
Procedure:
A simple apparatus for steam distillation is shown in
Fig.1.0.
It consists of distillation flask contains the liquid to be steam
distilled; it is fitted with the
'splash-head, which prevents the cary-over of the contents of the distillation flask into the
receiver. Place the solution or mixture of solid with a little water in distillation flask. Then
assemble the apparatus properly. Pass the steam into distillation flask. Start
heating the
distillation flask slowly and continue the passage of steam until no appreciable amount of
water-insoluble material is received in receiver flask. Discontinue the passage of steam when
the distillate is completely recovered in flask.

4/Unit-II
 Safety tube Condensed
water vapour
Steam
generator Water outlet
Stand
bath Water inlet
Water

Distilled liquid
ICompound water
to he distilled

Fig. 1:Steam Distillation Assembly

A Practical Book of Pharmaceutical Organic Chemistry-lI /5

 Synthesis of Few
UNITV Organic Compounds

A) Synthesis of Benzanilide

H,N -Cl NH-C-CHs

NaOH
+H,O+ NaCl

Anilline Benzoyl Chloride Benzanilide

Principle:
Insertion of Benzoyl moiety with replacement ofhydrogen atom from OH, NH, is called
Benzoylation reaction. Benzanilide is synthesized by Schotten Baumann Reaction. In this
reaction, Benzoylation of amines are caried out using Benzoyl chloride, in presenceofaqueous
Sodium hydroxide. Benzoylation proceeds smoothly and sparingly soluble derivative usually
separates as a solid. Excess Benzoyl chloride is hydrolyzed by Sodium hydroxide as Sodium
benzoate and Sodium chloride, which remains in the solution

H,C-NH, + H,C C Cl -H+

H,C-NH-C-Ci

Aniline Benzoyl chloride

-c
H,C-NH- H,C-NH-C-CH,
Benzanilide
Procedure: Place 2.6 g(2.5 ml) ofAniline and 25 ml
of 10% Sodium
solution in 100 ml conical flask. Then add 4.3 hydroxide (AqueousS)
g (3.5 ml) of Benzoyl chloride.
10minutes. Heat is evolved during reaction. When Stopper
flask and shake the
vigorously
is complete (i.e. when the
for the reaction
white fumes cease), filter off the
crude wash with water and
recrystallise fromrectified spirit.
Yield is 4.4gand
Melting point 162°C.
34/Unit-V
B) Synthesis of Phenyl Benzoate:

OH -CI

NaOH R
-0-¢

Phenol Benzoyl Chloride Phenyl Benzoate

Principle: This synthesis is based on Schotten Baumann Reaction. Phenols are benzo-
ylated in presence of aqueous Sodium hydroxide.

OH
=0 H -0

-C -HC

Phenol Benzoyl Chloride Phenyl Benzoate

Procedure:
Dissolve 2.5 gm ofphenol in 38ml of 10% Sodium hydroxide in250 ml lodine flask. To it
add 4.5 mlof Benzoyl chloride and shake the mixture vigorously for 10 minutes. (Caution).
Filter offthe Phenylbenzoate which separates and wash with cold water.
Recrystallisethe crude using rectified spirit (colourless crystals).
Report the yield
Yield(4.1 g); Melting Point. 68° C.

A Practical Book of Pharmaceutical Organic Chemistry-II/35

 C) Synthesis of Acetanilide

O
NH,
HN -CH,
HC C Conc.HCI1
HC -OH

HC-

Aniline Acetic Anhydride Acetanilide

Principle
Acetanilide is synthesized from Aniline by acetylating it with Acetic anhydride in
presence of
glacial Acetic acid. Aniline or Phenyl amine is a primary amine and basic in nature. Acetic
anhydride, an anhydride of Acetic acid, acts as a source of Acyl group. Aniline reacts with
Acetic anhydride to form Acetanilide by nucleophilic substitution reaction and
the reaction is
called Acetylation. In this reaction,Aniline acts as a
nucleophile and Acetyl group (-CHCO
from Acetic anhydride act as an electrophile. Here, the hydrogen atom of-NH, group is
replaced by the acety l group.
Mechanism:

NH,
H,C H,C
H,C-
H,C-

NH-C-CH, + CH,COO

Procedure:
Take 125 ml of Water in 250 ml beaker and add 4.6 ml
of conc. HCI to it. To this add 5.1g
(5.0 ml) ofAniline and stir until it completely passes into solution. Add 1 gofcharcoal powder
ifthe solution is coloured and wamm the mixture at 50°C with stirring for 5 minutes. Filter with
fluted filter paper. Add 6.4 ml (6.9g) of Acetic anhydride to the resulting solution and stir.
Prepare a solution ofSodium acetate (8.3g) in 25 ml water. Add this solution to above reaction
mixture with stirring and cool in ice bath.
36/Unit-V
 Cilter the crude acetanihde using vacuum
pump and wash well with water. Recrystallize from
125 ml of boiling water with 3 ml of Methylated spirit.
Yield: 4.99 g M.P: 144°C

D) Synthesis of 2,4,6-Tribromo Aniline

NH,
NH
Br Br
3Br2 3HBr2

Br

Aniline 2,4,6-Tribromo aniline

Principle:
Aniline undergoes electrophilic substitution with Bromine, even in cold. The bromine atoms
enter at the two ortho positions and para positions with the formation of2, 4, 6,-tribromoaniline.
Here presence of bromine atoms in tribromoanilline, reduces the basic properties of amino
group, salt even with strong acids are almost completely hydrolyzed in presence of water.
When Br' attacks the ring as electrophile, it generates pentavalent carbon atom, one ofthe
double bond in the ring breaks, as H* leaves, to give stability and bond re-forms, NH, group
is releasing group which facilitates ortho and para substitution.

Mechanism:

A Practical Book of Pharmaceutical Organic Chemistry-1 /37

 M) Synthesis of Dibenzalacetone from Benzaldehyde

O
CHCOCH
--H NaOH or A.q.EtOH
-CH=CH-Ë-CH=CH

Benzaldehyde
Principle:
When an ethanolic solution containing acetone and its two equivalents of Benzaldehyde is
made alkaline with sodium hydroxide, rapid condensation occurs with the formation of
Dibenzalacetone. This is a particular example of Claisen reaction. Claisen showed that
aldehydes under the influence of Sodium hydroxide condense with, (i) another aldehydes, (ii)
or ketones, with the elimination ofwater.
Thus, Benzaldehyde condenses with () acetaldehyde
to give Cinnamic
aldehyde, and with (ii) equivalent of acetone to give
(mono) benzal-acetone
Procedure:

OH
H,C-C- CH-H Step1H,C-t- H,-
Step 2
O

H,C--CH- H,O
H QH
H
Step 3
H,C-
H OH
OH Step 4
H,C-C Step 5

H
-H.O -C-cH=CH-
H
A H
Repeat Step 1 to 5
=CH--c=C-
Again H

Dibenzalacetone
48/Unit-V
Procedure:

Mix about 10 ml (10.4gm) of benzaldehyde and 4 ml of pure acetone with 10ml of

spirit in a conical flask. 2ml of 10% aqueons sodium hydroxide is diluted with 8 ml of
methylated
water
and this dilute alkali solution is added to the former solution. The mixture is shaken
vigorously
in stoppered flask for about 10 minutes ( releasing the pressure from time to time
ifnecessary)
and then allowed to stand for 30 minutes with occasional stirring and finally cool in ice-bath.
During shaking, the dibenzal-acetone separates at first as a fine emulsion which then rapidly
forms pale yellow crystals. Filter off the product at pump. Wash with ice- cold water. Dry the
solid at room temperature upon a filter paper to constant weight. Recrystallise in hot ethyl
acetate (2.5 ml per gram) or in hot rectified spirit. Yield: 80%, M.P. 122°C.

nlDonlr of RhormocauticalOrganic Chemistry-I1/49

 ) Synthesis of benzoic acid from alkyl benzoate
Reaction
HSO cOOH
A ONa +C.H,OH-
C-oc,H+ NaOH

Principle: Esters are hydrolyzed either by an acid or base. Alkaline hydrolysis of ester is
irreversible, which is also called Saponification. The common mechanism is: The alkaline
hydrolysis, which occurs through a nucleophilic acyl substitution. Here Ethyl benzoate on
hydrolysis with Sodium hydroxide gives Benzoic acid and Ethyl alcohol, where OH ion of
Sodium hydroxide acts as a nucleophile.
Mechanism:

CH-C-Oc.H,CH,--Oc.H,CHcoOHCH,o
HO OH
H
HHExchange HC-
Exchange H.C-c-0+ OCH, CHCOOH+C,H,

Procedure: Take 2 ml of Ethyl benzoate and add Sodium hydroxide solution (15 ml, 10%).
Reflux the mixture for 30 minutes in water bath (90-100°C) till the ester layer disappears.
Then cool the solution and acidify with HCl. Cool the resultant acidified solution in an ice
bath.
Filtertheseparated Benzoic acid precipitate and recrystallise in hot water. Yield: 1.2 g
Melting Point 122°C.

44/Unit-V
 nSvnthesis of 1-Phenyl Azo 2- Napthol From Aniline

OH
CI
NH +H,0
+ NaNO, NEN Y N= N
-HCI
-2H,O Bnapthol
- NaCl

Anilline
1-phenyl azo 2- napthol

Procedure:
Dissolve 4 gor 3.92 ml of aniline in 12.8 of concentrated Hydrochloric acid in 250 ml of
beaker and dilute it with 12.8 ml of distilled water. cool in ice bath with
frequent stirring till it
attained a temperature below 5°C. In another beaker, dissolve 3.2 g of
ml of water and chill the solution in ice bath
sodium nitrate in 15
(0-5°C), add sodium ntrite solution (2) to the
aniline solution (1) in small lots (2 ml), at a time in intervals with
vigorous stirring, with a glass
rod taking care that the temperature of reaction mixture must not exceed
beyond 5"C at any
cost. After the complete addition ofsodium nitrite
solution, it is required to test the reaction
mixture for the presence of free nitrite by taking out a drop ofit and
immediately placing it on
potassium iodide starch paper that will distinctly tum blue in the presence of free nitrous acid.
Dissolve 6.24 gof beta naphthol separately in 250 ml beaker in 40 ml
solution and cool this solution in ice bath. Slowly add the cold diazoniuum salt
ofSodium hydroxide
solution to the
beta naphthol solution with vigorous stirring,
Special care must be taken to not allow the
reaction mixture raised beyond 5°C. Ifrequired, add crushed ice while the
coupling reaction
proceed. Ared colour develops and crystal ofcrude phenylazo- beta napthol separates out.
Allow the reaction mixture to stand for 30-40 minutes with
stirring to complete the reaction.
Filter the red product in Buchner funnel using suction, and wash with ice cold
water, drain the
water.

Crude Yield 90% Melting Point. 129-130°C

A Practical Book of Pharmaceutical Organic

Chemistry-I/45