Experiments

29
Chemistry - I
Dxperimental Medicinal

on water bath forabout 2 hrs.

Refluxthe reactionmixture
of water and
reactionmixture into125
mL
Cool to room temperatureand pour the
mix carefully. an insoluble
under suctionto remove
to stand for 15 minutes and filter
Allow
by-product. acid, cool in
acidic with concentrated hydrochloric
Render the s
filtrate trongly
suction.
offprecipitatedproductunder
filter
ice-water and immediately spirit.
from 20-30mL of industrial
can be recrystallízed
The crude product
Recrystallization:
Theoreticalyield:2.8g (52%)
297-298°C.
Melting point:
Physicalconstant:
CalculationTable:
Actual Quantity Actual Molar
Sr. Reactant Molecular
Moles taken Ratio
No. Weight (g) taken (Unit)
0.03 1
1 Benzil 210 5.3

60 3 0.05 1.98
2. Urea

75 mL (density 50.97
3 1.29
Ethanol 46
= 0.789g/cm)

4 Sodium Hydroxide 40 15 mL
PracticalYieldx 100
The percentyield =
Calculation: Theoretical
Yield
Result:
1 The percentageyieldof phenytoin was found to be ......%.
w/w.
2. Physicalconstant:
3. Appearance:

Experiment 8:PreparationofPhenothiazine

Aim: To prepare phenothiazine

from diphenylamine.
Requirement:

absolutealcohol.
Chemicals: Diphenylamine,sulphur,aluminium chloride,

Apparatus:Beaker,round bottom flask,condenser,Buchner funnel, beaker, glass rod.

measuring cylinder,
filter
paper,thermometer.
Principle:

nenothiazineisprepared by fusingdiphenylamine with sulphur with rapidevolutionof

hydrogen sulphidein alsobe used,
presence of aluminium chloride.Calcium chloridecan
however yieldobtained
is less.
 Erperimental MediclinalChemistry -I B0
Experiments

Reaction:

S,AICI,

Dlphenylamine Phenothiazine

Procedure:

Take 22 g (0.13mole) of diphenylamine, 8.2 g (0.26 mole) of sulphurand 3.2g

(0.03mole) of anhydrous aluminium chloridein a beaker and melt them togetheron
a heatingmantle.

The reactionmay attaintemperatureup to 140-150°C

with the rapid evolutionof
hydrogen sulphide.

By loweringthetemperature,a few degreesthe reactioncan be slackened.

When the reactionis settled,
raisetemperatureto 160°Cforsome time.

Ground up the melt, when cool,and extractfirstwith water and then with dilute
alcohol.
The residueconsistsofalmost pure phenothiazine
as yellowishleaflets.
Note:Releaseof HS can be checked by holdingmoist lead acetate paper at the top of the
condenser.Blackeningof paper indicates
releaseofHS.
Recrystallization:
The crudeproduct can be recrystallized
from absolutealcohol.
Theoreticalyield:
19.8g (90% w/w)
Physicalconstant:
Meltingpoint:179-180°C.
Calculation
Table:
Sr. Reactant Molecular Actual Quantity Actual Molar
No. Weight (g) taken (Unit) Moles taken Ratio
1. Diphenylamine 169 22 g 0.13 1
2 Sulphur 32 8.2g 0.26 2

3. Aluminium Chloride
133 3.2g 0.03 0.22
(anhydrous)

Practical Yield x 100

Calculation:
The percentyield=
TheoreticalYield
Result:
1. The percentageyieldof phenothiazine was found to be % w/w.
2. Physicalconstant:
3. Appearance:
 Experiment14:Aspirin
Aim: To performassay'of
aspirin.
Requirements:
Chemicals: Aspirin,sodium hydroxide,ethanol (95%),concentratedhydrochloricacid,
phenolred indicatorsolution.
Apparatus:Beaker,conical flask.pipette, pipettefiller,
burette,
measuring cylinder,
burette
stand and holder,
glassfunnel, white tile,
bench mat.

Principle:
1 The assay of aspirin(.e.acetyl acid)is carriedout by
salicylic aqueous acid base
titration
which is based on simpleneutralization
reaction.
2. Itis a back type oftitration.
3. The assay ofaspirin(back titration)
can be explainedstepwiseas givenbelow:

Steps:
StepI:
Alkalinehydrolysisof aspirinwith excess NaOH.

A known excess of standard alkalii.e.NaOH is added to a weighed amount of the

aspirin.
oCOCH, OH
2NaOH +
CH,COONa + H,0
cOOH COONa
Aspirin Sodium hydroxide Sodium salicylate

of unreactedNaOH
Step : Back titration :
After the reaction (.e.alkalinehydrolysisof aspirin)is completed, the excess
ie.remaining NaOH (unreacted)is back titratedwith a standardsolutionof HCI using
This givesburettereading 1.
phenol red as an indicator.
 Bxperimental Medicinal Chemistry
-1 53 Experiments

Step II: Blank determination:

using the same procedure,only omitting the sample of aspirin.
A biank is run parallel

2.
This givesburettereading

Step TV: Differencein readingdetermination:

The amount of the titrantrequiredto reactwith the sample (analyte)is subtracted from
the main
reading 2) (Burette
ofthe titranti.e.(Burette
volume -reading 1).

Requirements:
Theory:
(A)Aspirin(Analyte):
1 Molecularstructure:
COOH
CH,

2. Molecularformula:CsH;O4.

3. Molecularweight:180.16.
acid,2-acetoxybenzoic acid.
4. Chemical names: Acetylsalicylic
5. Melting point:135°c.
It is white,crystalline,
Aspirin is an acetylderivativeof salicylicacid.
6. Description:
weakly acidicsubstance and odourless.

analgesic,anti-inflammatory,antipyreticand is
7. Pharmacological activity:
Aspirin is

an inhibitorof plateletaggregation. It inhibitsfatty acid cyclo-oxygenaseby

effectsof aspirin
acetylationof the activesiteof enzyme and the pharmacological
are products including
due to the inhibitionof the formation of cyclo-oxygenase
thromboxanes and prostacyclin.
prostglandins,

8. Uses:

0 Aspirinis used to treatpain, and reduce feveror inflammation.

(G) It is sometimes used to treator preventheart attacks,strokes,and chest pain

(angina).

Gi)Aspirinshould be used forcardiovascularconditionsonly under the supervision

of a doctor.

9. Storage:Store protectedfrom moisture at a temperature not exceeding 30°C.

Moisture can hydrolyze aspirintherefore
always kept in dry form.

Refer Assay of Chlorpromazine

(B) Potassium Hydrogen Phthalate (Primary standard):
Hydrochloride.
 Medicinal Chemistry
-I 54 Experiments

Bxperimental

()Sodium carbonate (Primarystandard):

1. Molecularstructure: :0:
Na

2. Molecular formula: Na,CO;

3. Molecular weight: 106.01
4. Equivalentweight: 53
5. Chemical names:Soda ash;carbonicacid;disodium salt;
disodium carbonate.
6. It is a diacidicbase: hence itsequivalentweight is half of its molecular weight
53.0.
i.e.

7. Melting point:851°C.
8. Solubility:
Solublein water,and insolublein alcohol,ethanol.
9. Description:Sodium carbonate is an inorganic salt.It is white, crystallineand
hygroscopic,odourlesspowder.
10.Uses:
) As a primary standardforacid-basetitrations.
(ii)Itisa 'washing soda'.

(i)
In toothpastes,
as a foaming agent and an abrasive.

(v)Itis a food additiveused as an acidity anti-cakingagent,raisingagent,

regulator,
and stabilizer.

()In manufacture of glass,as a pH regulatorto maintain stablealkalineconditions,

as an electrolyte
in chemistry.

(vi)Sodium carbonate,in a solutionwith NaC., may be used forcleaningsilver.

(D)Sodium Hydroxide (Titrant):

ReferAssay ofIbuprofen.

(E) Hydrochloric
acid (Titrant):

1. Molecular formula:HCI
2. Molecular weight:36.46
3. Equivalentweight:36.46
4. Itis a monoprotic acid;hence itsequivalentweight is same as its molecularweight
36.46.
i.e.

5. Chemical names: Spirits

of salt,
acidumsalis(saltacid),muriaticacid.

6. Description:
Hydrochloricacid is a strong inorganicacid. It is a clear,
colorless,
fuming liquid;having pungent odour.It is highly corrosive.Itis found
naturallyin
Oastric
acidwhich acidifies
stomach contentsto a pH of l-2.
 IMedicinal Chemistry -I 55 Experiments
Dperimental
issues:
7. Safety
o ConcentratedHCI (fuming HCI) forms acidicmists.Both the mist and thesolution
have a corrosiveeffecton human tissue, with potentialto damage respiratory
organs,eyes,skin,and intestines
irreversibly.
G) Personal protectiveequipments such as rubber or PVC gloves,protective eye
goggles, clothingand shoesare used to minimize risks.
and chemical-resistant

Gin The standard first

aid on the skin is,
measure if acid spills to remove and isolate
contaminated clothingand shoes.Immediately flushwith running water for at
least20 minutes.

Phenol red solution(Indicator):

1. Molecularstructure:
HO OH

2. Molecular formula:CgH4O;S.

3. Molecular weight:354.38
(PSP)
4. Chemical name: Phenolsulfonphthalein
and 2.9g/L in ethanol.
5. Solubility: 0.77 grams per litre(g/L)in water
red is
is stablein air. Phenol
exists as a red crystalthat
6. Description: Phenol red
red is 6.8 to 8.2.Phenol
as a pH indicator. The pH range of phenol
commonly used
6.8 and pink above pH 8.2.
red is yellow below pH a
This compound actually changes color through
7. Mechanism of color change:
very acidic
In crystallineform, and in solution under
called ionization.
process ion with the sulfategroup nega
pH), phenol red exists as a zwitter
conditions (low
proton and is orange
and the ketone group carrying an additional
tively charged, the ketone group islost,
increased (pKa = 1.2),the proton from
pH pH (pka = 7.7),
red. If the is

yellow negatively charged ion. At stillhigher

resulting in the
loses itsproton, resultingin thered ion.
hydroxide group
the phenol's

(G)Methyl red solution

(Indicator):
1. Molecular structure:

OOH
H,C

H,
 -I 56 Experiments
Medicinal Chemistry
Bperimental

2. Molecular formula:CsHysN,O,.
3. Molecular weight: 269.30.
acid.
4. TUPAC: 2-(4-Dimethylaminophenylazo)-benzoic
5. Melting point:182°C.
insolublein
Solublein ethanol,practically
6. Solubility: water.

7. Description: Itis one ofthe azo dyes.Itis a

Dark red powder or violetcrystals. pH

indicator.
8. pH range:4.2 to 6.3.
9. Colourchange:Red (inacidicmedium) to Yellow (inbasic medium).
10.Mechanism of colorchange: Itis based on Resonance theory'.The above color
change is associatedwith a rearrangement involving azo link.

ofReagents/Solutions:
Preparations

(a)0.5 M Hydrochloricacid: It is prepared by diluting42.5 mL of concentrated

hydrochloricacid to 1000 mL with distilled
water.

(b)0.5M Sodium hydroxide:It is prepared by dissolving20 g of NaOH to 1000 m

with distilled
water.

(c) Phenol red solution:Dissolve0.1 g of phenol red in 2.82 mL of 0.1 M sodium

hydroxideand 20 mL of ethanol (95%). water to produce
Add sufficient 100mL.
(d)Methyl red solution:
Dissolve50 mg of methyl red in a mixture of1.86mL of0.1M
sodium hydroxide and 50 mL of ethanol (95%).Add sufficient
water to produce
100 mL.

Standardization
Procedure:

of HCI:
(A)Standardization

1. Weigh accuratelyabout 0.75g of anhydrous sodium carbonate(which is previously

heated atabout270°C for1 hour) in an Erlenmeyerflask.

2. Dissolveitin 100 mL of distilledwater.

3. Add 0.1ml of methyl red solutionas indicator.

4. Add the 0.5 M hydrochloricacid slowlyfrom the burette,with constantstirring,

until

the solutionbecomes faintlypink.

5. Heat the solutionto boiling,cool and continue the titration.

6. Heat again to boiland titratefurtheras necessary until the faint pink coloris no
longeraffectedby continued boiling.
 I Medicinal Chemistry I
Byperimental Experlments
Table:
Observation
Sr.No. BuretteReading (mL) of HCI
Initial Final Mean
1 0.0 m A mL
A+ B+C
2 A mL B mL 3

3 B mL CmL -X mL

Reaction:

2HCI + Na,CO, 2NaCI + HCO, H,O + co

Sodium Salt Carbonic Water
Hydrochloric
carbonate acid
acid

FactorCalculation:
2 moles of HCI 1 mole ofsodium carbonate

2000 mL of 1 M HCI 106g of Na,CO,

1000 mL of 1 M HCI 53 g of Na,CO

1 mL of 0.5M HCI = 0.02649g of Na,CO,

Factor:1 mL of0.5M HCI = 0.02649g of sodium carbonate

for
lbuprofenand make necessay correction
(B)Standardizationof NaOH: Referassoy f

05 M NaOH.

Factor:1 mL of0.5 M NaOH = 0.1021g ofCgH,KO,

ASSAY OF ASPRIN:

1. Weigh accuratelyabout 1.5g aspirin.

2. Dissolveitin 15 mL of ethanol (95%).

3. Add 50 ml of 0.5 M sodium hydroxide.

4. Boil gentlyfor 10 minutes,and then cool it.

5. Add few drops of phenol red solution(asindicator).

alkali)with 0.5 M
6. Titrate the above solution (i.e.the excess or unconsumed
acid to get end pointas permanent faintpink color.
hydrochloric

1. Repeat the same procedure without taking the substance under examination

This
without aspirin:
(i.e. is called'blankdetermination').

8. The differencebetween represents the amount of sodium hydroxide

the titrations

required.
 Experiment 13:
lbuprofen

Aim: To perform assayy ofibuprofen.

Requirermernts:
0.1M sodium
demicals:Ibuprofen, ethanol(95%),phenolphthalein.
hydroxide,
Aenaratus:Beaker,coniclflask,
pipette,
pipettefiller,
burette,measuring cylinder,
burette
glassfunnel,
holder, bench mat.
white tile,
standand

Principle:
. This assay determinesthe contentof ibuprofenby acid-basetitration ibuprofen
i.e.
as an indicator.
againstNaOH
(acid)istitrated using phenolphthalein
(base)

Theory:

A) lbuprofen(Analyte):

1 Molecularstructure:
CH,
CH, COOH

H,C
C3tisO2
2. Molecularformula:

3. Molecularweight:206.3
acid.
4. Chemical names: Ibuprofenis (RS)-2-(4-isobutylphenypropionic
5. Meltingpoint:
75-78°C with
crystals
powder or colorless
Itis a white or almost white crystalline
*vescription:
slight
odour.
solublein water.
slightly
Itisvery solublein alcoholand
solubility:
 - I Experiments
Experimental Medicinal Chemistry

8. Pharmacologicalactivity:
inhibitsthe activityofCOX-I and COx-n
(0 Ibuprofenis a propionicacid derivate,
resulting in a decreased formation of precursors of prostaglandinsand
thrombOxanes.

()t is non-steroidalanti-inflammatory drug (NSAID) with anti-inflammatory,

analgesic,and antipyreticeffects.

(B)Potassium Hydrogen Phthalate(Primary standard):

ReferAssay of Chlorpromazine
Hydrochloride.

()Sodium Hydroxide (Titrant):

1. Molecularformula:NaOH
2. Molecularweight:40
3. Equivalentweight:40
4. Molecularweight and Equivalentweight aresame as it is a mono-acidic base i.e.
40.
5. Chemical names: Causticsoda (Itis a highlycausticmetallicbasse),Lye.
6. Melting point:121-124°C.
7. Solubility:
Soluble in water,ethanol and methanol. It is deliquescent and readily
absorbs moistureand carbon dioxidein air.
8. Description:White, waxy, opaque crystallinesolid,usually in the form of semi
sphericalpellets,
flakes,granules and
it is odourless.It is
hygroscopic in nature.
Always containsa definiteamount of sodium carbonate and
it
water. Therefore,
solutionsof NaOH will need to be
standardized.Itis usuallydispensed as semi
spherical
pellets.
9. Safetyissues:
) Specialcare is
required to prepare a solution of NaOH
in water because
considerableheat is liberatedby the
exothermic reaction.The solutionmay
splatteror boil.

(i)Like other corrosiveacids

and alkalis, drops of sodium hydroxide
solutionscan
decompose proteinsand lipids
skin, eyes or other living
hydrolysis tissuesvia amide
and esterhydrolysis
which consequently causes
may induce chemical burns and
permanent blindness, if itcontactseyes.
(i) Protectiveequipments such as rubber
gloves,safetyclothingand eye
should always be used protection
when handling sodium
hydroxide solution.
(iv)The standard
firstaid measure
foralkalispills on the skin is
quantitiesofwater forat washing with large
least10-15minutes.
 -T Experiments
EperimentalMedicinal Chemistry

(o PhenolphthaleinSolution(Indicator):
1 Molecularstructure:

H
HO"

2. Molecularformula:CzoH4O4
3. Molecularweight:318.3
260°C.
4. Meltingpoint: in basic solutionsor in ethanol
or
5.Solubility:Insolublein pure water, but soluble
acetone.
a phenol
3-bis(4-hydroxyphenyl)phthalide.It is
6. Chemistry: Phenolphthaleinis 3,
derivativeofphthalic anhydride.
and odourless powder. It is a very
7. Description: A white, crystallineor amorphous It
involving weak acidsand strong bases.
Itis used in titrations
popularpH indicator.
turnscolorlessin acidicsolutions and pink in basic solutions.

8. pH range:8.3to 10.0.
is colorlessbelow pH 8 and pink above pH 9.6.
9. Mechanism of colorchange:
is a weak acid,it contains positivelycharged hydrogen ions,
Phenolphthaleinitself
those ionsare removed. As a whole,
but when it is exposed to an alkalinesolution,
but its ion is naturallybright
the phenolphthaleinmolecule is naturally colorless,
pink. When the hydrogen ions are removed from the solution,the colorless
the
phenolphthaleinmolecule ionizesand changes to the pink color.The stronger
alkalinesolutionis,the more hydrogen ions are removed and
the darker the pink
colorof phenolphthalein.
OH OH

HO
-P-

pH 8 (Colourless) pH 9.6(Pink)

Preparationof Reagents/Solutions:
(a)0.1M Sodium hydroxide:It may be prepared by dissolving 4 g of sodium
water toproduce1000mL
carbon dioxide-free
hydroxide in sufficient

(b)Phenolphthaleinsolution:
A 1.0%W/v solution
of phenolphthaleinin ethanol(95%).
 Experimental MedicinalChemistry - I 50 Experiments

StandardizationofNaOH:

1 Weigh accurately about .5

g of potassium hydrogen phthalate (should
he

2 hours).
previouslypowdered and dried at 120°Cfor
it in 25 mL
2. Dissolve water
ofcarbon dioxide-free in a flask.

3. Add 0.1mL ofphenolphthalein solutionas indicator.

4. with 0.1M NaOH solution.

Titratethisabove solution

5. The end pointis a permanent pink color.

Note:
1 StoreNaOH suitablestoppersto protectfrom
solutionsin bottleshaving well-fitted
atmospheric CO
ofsodium hydroxidemust be regularly.
re-standardized
2. The standard solutions
ObservationTable:
Sr. BuretteReading (mL) of NaOH
No. Initial Final Mean

1. 0.0mL A mL
A+ B+C
2. A mL B mL

B mL C mL =X mL
3

Reaction:
COOK COOK
NaOH +
H,0
COOH cOONa

Potassiumhydrogen Sodium potassiumphthalate

phthalate (KHP)

FactorCalculation:

1 mole of NaOH = 1 mole of potassium hydrogen phthalate

1000 mL of 1 M NaOH = 204.2g of C&HsKO4

1 mL of 1 M NaOH 0.2042g of CgHsKO4

1 ml of0.1M NaOH = 0.02042g of CgHKO4
1 mL of0.1M sodium hydroxide is equivalentto0.02042g ofCgHsO4.
ASSAY OF IBUPROFEN:
Procedure:
Weigh accuratelyabout0.4g ofIbuprofenand dissolvein 100 ml of ethanol(95%).
The resultantsolution was titratedwith 0.1M sodium hydroxide
solutionusing
phenolphthaleinas an indicator.

Carry out blank determination as well.

 Experimental Medicinal
Chemistry -[ Experiments

Table:
Observation
BuretteReading (mL) of NaOH
Sr.No.
Final Mean
Initial

0.0mL A mL A+ B+C
1
3
B mL
A mL
2. =X mL
C mL
B mL
3.

Reaction: CH,
CHg ONa
CH,
+ Hy0
+ NaOH
H,C
H,
FactorCalculation:
1mole of Ibuprofen
1 mole ofNaOH
206.3 gm oflbuprofen
1000mL of 1 M NaOH

1 mL of 1 M NaOH 0.206.3gm of Ibuprofen

1 ml of 0.1M NaOH = 0.02063gm ofIbuprofen

1mL of 0.1M NaOH = 0.02063g of CHHs02

Factor:

of 0.1M sodium hydroxide = 0.02063g ofIbuprofen

1 mL

Calculations:

Step : (Normality of NaOH)

lml of0.1M NaOH = 0.02042g of CgHsKO4

mLmeon burete reoding Of y M NaOH 0.5g of CgHsKO,

0.5 x 1 x1
y = (X ml) x (0.02042)

y =

Step I:(Amount ofIbuprofen)

1 mL of0.1M NaOH = 0.02063g ofIbuprofen

X mlBurette reading of assa) of y M NaOH = Ag ofIbuprofen

X mL xy x 0.02063g
Ag = 1x0.1
 52 Experiments
Chemistry -I
Experimental Medicinal

StepM: (% Purity)
For 0.4 g of Ibuprofen 100%

Ag ofIbuprofen P%
(100) x (A)
Hence,
P = 0.4

98.5% and not more than 101.0%of CuH0,

Limit:Ibuprofencontains not less than
on thedried basis.
calculated
Result:

1. The molarityof Sodium hydroxidesolution

0.1M) was found to be
(prepared, .....
ofIbuprofenwas found to be .....%w/v
2. The percentpurityofgivensample
 perimental Medicinal Chemtstry ! 39 Experiments

Factor:
1mL of0.1
M HCIO, 0.03553g ofChlorpromazinehydrochloride
Calculations:
Step I(Normalityof HCIO)
1ml of0.1M HCIO,s0.02042g ofpotassium hydrogen phthalate
X mLsurettereoding stondardizotionyOf y M HCIO, 0.35g of CHsKO4
of
0.35 x 1 x 0.1
y =X mL) x (0.02042)

y = .....
M

Step (Amount of Chlorpromazine HC)

1 mL of0.1M HCIO, = 0.03553 g ofChlorpromazineHydrochloride

assay)Of y M HCIO4 = Ag ofChlorpromazineHydrochloride

X mLBurettereading
of
X mL xy x 0.03553g
Ag = 1x 0.1

Step (% Purity)
100%
For 0.6g of Chlorpromazine Hydrochloride
P%
Ag of ChlorpromazineHydrochloride
(A)
P = (100)
X06
Hence,
than 101.0% of
contains not less than 99.0% and not more
Limit:Chlorpromazine HCI
on the dried basis
.
GH,gCIN:S- HC, calculated
Result:
to be....M.
acid solution(prepared,0.1M) was found
1 The molarity of perchloric
to
of Chlorpromazine Hydrochloride was found
purityof given sample
2. The percent
be ...%w/v

Experiment 11:
Phenobarbitone

of Phenobarbitone.
Aim: To perform assay
Requirements: silver
solution,
ethanol,sodium hydroxide,thymolphthalein
Chemicals: Phenobarbitone,
pyridine.
nitrate, burette
measuring cylinder,
burette,
filler,
pipette
conical flask,pipette,
Apparatus:Beaker, bench mat.
g
holder, lass funnel,white tile,
standand
 Experiments
MedicinalChemistry
-I
Experimental

Principle: titrationusing sodium hydroxide

estimated by non-aqueous
Phenobarbitonecan
be
1
as titrant.
aqueous media, a suitable non-aqueous
as weak acid and
iseasilysoluble
a
Since, it functions
2.
agente.g.DMF or Pyridinein which it
solventis used as levelling iune
in quantitativemanner as itssilversalt
out from pyridine
3. Itcan be precipitated
additionof silver nitrate. Nau
HNO, can be titratedwith
containingdisplaced
4. The supernatant pyridine,
solution.

Theory:

(A)Phenobarbitone(Analyte):
1. Molecular structure:

2. Molecular formula:C2H12N,0,
3. Molecular weight:232.2
acid.
barbituric
4. Chemical name: 5-ethyl-5-phenyl
5. Melting point:174-178°C
Itis colorless
6. Description: powder and odourless.
crystalsora white,crystalline

It is sparinglysoluble in water and benzene, freelysolublein alcohol,

7. Solubility:
chloroformand ether.Itis alsosolublein alkalihydroxidesand carbonates.
8. Pharmacological activity:
) Phenobarbitalbinds to and activatesthe gamma-aminobutyric acid (GABA)-A
receptor,therebymimicking the inhibitoryactionsofGABA in the brain.

G) Phenobarbital is a long-actingbarbituricacid derivativewith antipsychotic

property.
(i)
Itisused as anticonvulsants,
centralnervous system depressantsand sedatves.
(B) Benzoicacid (Analyte):
1. Molecularstructure:
COOH
 Experiments
Chemistry - I 41
Experimental Medicinal

2. Molecularformula:C,H,O,
3. Molecularweight:122.1
122.4°c
4. Melting point:
solidwith faintand pleasant odour.
Benzoicacid is white crystalline
5. Description:
6. Uses:
(0) Itssaltis used as food preservative.

by bacteria.
(i)Ithelpsto prevent infectioncaused
and
acid is used to treatskin irritation
with salicylic
(ii)Benzoicacid in combination
or eczema.
insectbites,fungal infections
inflammation caused by burns,

()Thymolphthalein (Indicator):
1. Molecular structure:

CH,
H

H,c
CH.
HO"
H,C CH,

2. Molecular formla: C2sHao04

3. Molecular weight:430.5
(4-hydroxy-2-methyl-5-propane-2-ylphenyl)-2-benzofur
4. Chemical name:3,3-bis
1-one.
range is around pH 9.3-10.5.
Its transition
Itis acid-baseindicator.
5. Description:
and
this pH, it is colorless
above this,it is blue.
Below
and dimethyl formamide.
Itis solublein ethanol
6. Solubility:

Preparationof Reagents/Solutions:
in dimethyl
A 0.1%w/v solutionof thymolphthalein
solution:
(A)Thymolphthalein
formamide.
of pyridine.
8.5g of silvernitratein 100 mL
(B)Silvernitrate(8.5%w/v):
hydroxide in S mi of
sodium hydroxide: Dissolve4.2gof sodium
(C)0.1M ethanolic ml. Allow the
aldehyde-free ethanol to produce 1000
water and add sufficient the
bottlefor 24 hours.Then quicklydecant
tightly-stoppered
solutionto stand in a
container.
tightly-closed
a suitable,
clearsupernatantliquidinto
 Experiments
Chermistry -I
Bperimental Medicinal

STANDARDIZATION:
Standardizationof ethanolicsodium hydroxide:
a mixture of 30 mi of
g of benzoic acid, dissolvein
1 Weigh accuratelyabout 0.6
ethanol (95%)and 6 ml ofwater.
2 Titrate with the ethanolic sodium
as indicator.
thymolphthalein solution
hydroxide solution, using 0.2 mL f

Observation Table:
Sr. BuretteReading (mL ofEt.NaOH)
Final Mean
No. Initial

1. 0.0mL A mL A + B+ C

3
2. A mL B mL
= X mL
3. B mL C mL

Reaction:
COOH COOH

Factorcalculation:
|
Benzoic acid
NaOH

Sodium benzoate
H,0

1 mole of sodium hydroxide = 1 mole of benzoic acid

1000 mL of 1 M NaOH 122.1g of C,H5O,
1 mL of 1M NaOH 0.1221g
ofC,HsO2
1mL of0.1M NaOH = 0.01221g ofC,H502
ASSAY OF PHENOBARBITONE:
Procedure:

• Dissolve0.1g of Phenobarbitone in 5 ml of pyridineand add 10 mL of silvernitrate

(8.5%w/v) in pyridineadd 0.25mL of thymolphthaleinsolution.
Titratewith 0.1M ethanolicsodium hydroxide untila blue coloris obtained.
Carry out blank titration
as well.

• The differencebetween the titration

representsthe amount of sodium hydroxide
required.
ObservationTable:
Sr. Reading (mL of Et.NaOH)
Burette
No. Initial Final Mean
1 0.0mL A mL A + B + C
2. A mL B mL

3. B mL C mL =X mL
Experimental Medicinal Chemistry -I 43
Experimnents

Reaction:

OAg

NH + 2 AgNO, NH + 2 HNO,
silver
nitrate Nitrdeacld
Ägo

Phenobarbitone
2 HNO, + 2 NaOH2 NaNO3 + 2H,0

FactorCalculation:
2 HNO, = 2AgNO;
2 moles ofethanolicsodium hydroxide
sodium hydroxide
2 moles of ethanolic
lmole ofphenobarbitone
= 232.2g CHizO,N, 2HNO,
2000 mL of 1 M ethanolicsodium hydroxide
g CizH0,N, 1HNO,
hydroxide = 116.1
1000mL of 1 M ethanolicsodium
hydroxide = 11.61
g Czti;N
1000 mL of 0.1M ethanolicsodium

1mL0.1 M ethanolicsodium hydroxide 0.01161

g CazH2OaN2

Factor:
g Phenobarbitone
= 0.01161
1mL 0.1M ethanolicsodium hydroxide
Calculations:
NaOH)
Step I:(Normality of Et. g of Benzoic acid
(Et.NaOH)=0.01221
1 ml of 0.1M ethanolicsodium hydroxide
of y M Et.NaOH = 0.6g of C,H,0,
XmLBurette reoding ofstondardizotion)

0.6 x 1 x 0.1
y = (X mL) x (0.01221)

y = ....
M

Step : (Amount of Phenobarbitone

)
(Et.NaOH)= 0.01161g
Phenobarbitone
1 mL 0.1M ethanolicsodium hydroxide

X mLeurette reading of assay)Of y M Et.NaOHa Ag of Phenobarbitone

X mL xyx 0.01161lg
Ag = 1x0.1

Step : (% Purity)
100%
For 0.lg of Phenobarbitone
Phenobarbitone P%
Ag of

Hence,
(100)x (A)
P = 0.1
 - I Experiments
Experimental MedicinalChemistry

than 99.0% and not more than 101.0% t

Limit Phenobarbitone contains not less
CzN,O, calculatedon thedried basis.
Result:
0.1M) was found to
hydroxidesolution(prepared,
1 The molarityof ethanolicsodium
be ......M.

The percent purityofgiven sample ofPhenobarbitonewas found to be.....%w

2.

Experiment12:AtropineSulphate

Aim: To perform assay of atropinesulphate.

Requirements:
Chemicals: Atropine sulphate,perchloricacid, anhydrous glacial acetic acid, acetic
crystalviolet.
anhydride,potassium hydrogen phthalate,

Apparatus:Beaker, conical flask,pipette, burette,measuring cylinder,burette

pipettefiller,
glassfunnel,whitetile,
stand and holder, bench mat.

Principle:

Atropinesulphate can quantitativelybe determined by titration

in non-aqueous
solvents.

The totalalkaloidsof the atropine group are determined by titrationagainst

perchloric
acid in anhydrous aceticacid.

The method can be applied to tablets,suppositoriesand eye drops containing

atropine.

The endpoint is determined potentiometrically

or with crystalvioletas indicator.
Theory:

(A)Atropinesulphate(Analyte):
1. Molecular structure:
CH,

OH

2. Molecular formula:
(C,Hz3NOs)2.H,SOH,0
3.Molecular weight:6768