Vishnu Institute of Pharmaceutical Education & Research

Vishnupur, Narsapur 502313, Medal District

CERTIFICATE
Certified as a benefited record of practical work done by
Mr. /Miss
Regd. No.of ..B.Pharmacy / M.Pharmacy
in the Subject
.. during the year 2015-2016.
No. of Experiments done:

I/C LECTURER

PRINCIPAL
1

INDEX
Sino

NAME OF THE EXPERIMENT

DATE

PAGE
NO.

GRADE

SIGNATURE
OF LECTURER

Estimation of Paracetamol by UV-Visible

Spectroscopy
Aim: To estimate the percentage purity of paracetamol by calibration curve method.
Apparatus: Volumetric flask, pipette, curettes.
Chemicals: 0.1M NaOH, paracetamol pure drug and tablets.
Principle: Paracetamol is widely used as analgesic and anti-pyretic. Paracetmol is chemically 4hydroxy ace amide and can be estimate by UV-spectroscopy. The principle involved is the
absorption of radiation due to the presence of electrons in the paracetamol that can be raised to
highly energy levels by absorption of energy. The absorbance was measured at 257nm and the
calibration curve is constructed to determine the percentage purity of the compound it obeys
bears low in the range of 2.5-15g/ml.
Procedure: Preparation of standard dilutions:

100mg of paracetamol was weighed accurately and dissolved in 50ml of 0.1M NaOH and
then diluted it to 100ml with distilled water.
1ml of above solution was taken and diluted to 100ml with water.
From the above stock solution 6 standard dilutions are prepared by diluting 1ml, 2ml,
3ml, 4ml, 5ml, and 6ml with 0.1M NaOH to 10ml.
The concentrations that are obtained are 1/ml, 2, 3, 4, 5, 6g/ml and the absorbance was
measured by using 0.1M NaOH as blank.
The calibration curve was plotted between concentration and absorbance.

Assay:

20 tablets were taken and weighed accurately and the average weight the tablets were
calculated and made then into powder.
Equivalent weight of 0.15gms of paracetamol was weighed accurately and dissolved in
50ml of 0.1M NaOH and diluted it to 100ml with water.
Shake it for 5min and than filtered.
10ml of above filtered solution was taken and diluted to 10ml with distilled water.
From this 10ml was pipette out and 10ml of 0.1M NaOH was added to it and then diluted
to 100ml water and mixed well.
The absorbance of the solution was measured at 257nm against the blank solution.

Category: Analgesic.
Report: The percentage purity of paracetamol by calibration curve method was found to be
________

Estimation of Ibuprofen by UV-Visible

Spectroscopy
Aim: To estimate the percentage purity of Ibuprofen by calibration curve method.
Apparatus: Volumetric flask, pipette, curettes & beaker.
Chemicals: 0.1M NaOH, Ibuprofen pure drug, water & Ibuprofen tablets.
Principle: Chemically Ibuprofen is [4-(2-napthyl propel phenyl) prop ionic acid]. Ibuprofen is a
non-steroidal anti-inflammatory drug used for the treatment of arthritis fever and as an analgesic.
The principle involved in assay of Ibuprofen is the molecular absorption in the UV-range (200400nm) Arises from the transition involving the valence electrons and the absorption maxima is
at 264nm.
The standard calibration curve was plotted and the sample was estimated and the percentage
purity was determined.
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Procedure: Preparation of 0.1N NaOH: 4gms of NaOH was dissolved in 100ml of CO2 free
water. Which give 0.1N of the NaOH solution
Procedure for calibration curve:

10mg of the Ibuprofen was dissolved in 10ml of the 0.1N NaOH which gives the 1gm/ml
as a stock solution.
From the above solution, 2.5ml was taken into a 25ml volumetric flask and make up to
the mark using 0.1N NaOH which gives 100g/ml concentration.
From these solution 0.5ml, 5ml, 1.5ml, 2ml, 2.5ml was pipette out in 10ml volumetric
flask and finally the concentration was made up to the mark, which gives the
concentration of 5, 10, 15, 20, and 25g/ml.
The absorbance of the solution was measured at 264 nm using 0.1N NaOH as blank.

Assay:

20 tablets were weighed accurately and powdered.

100mg Equivalent weight of drug was taken and dissolved in 0.1N NaOH and final
volume was made up to 10ml using 0.1N NaOH.
From the color solution. 1ml was taken add and diluted up to 100ml in the volumetric
flask which gives 10g/ml.

Category: Non-steroidal, Anti-inflammatory drug.

Report: The percentage purity of Ibuprofen tablets by calibration curve method was found to
be _________.

Estimation of Nimisulide by UV-Visible

Spectroscopy
Aim: To calculate the percentage purity of Nimisulide by UV-Visible spectroscopy.
Apparatus: Volumetric flask, Curette, Pipette.
Chemicals: Nimisulide pure drug, Nimisulide tablets, 0.1N NaOH.
Principle: Nimisulide is a non-steroidal anti-inflammatory drug. The principle involved in the
assay of Nimisulide is the molecular absorption in the visible region that rises from the emerging
transitions which involves the valence electrons. The absorbance is measured at 393nm using
0.1N NaOH as blank. It obeys beers lamberts law in range of 10-35g/ml.
Procedure:
For Calibration Curve:4

100mg of pure drug was dissolved in 50ml of 0.1N NaOH solution and finally the
volume was made up to the mark with 0.1N NaOH which gives the concentration of
1mg/ml.
From the above stock solution 10ml was taken into 100ml volumetric flask to that 50ml
of 0.1N NaOH solution was added and the volume of the mark was made up with
distilled water.
0.5, 1.5, 2&2.5 of the above solution was taken into 10ml volumetric flask and is made
up to the mark with distilled water.
The concentrated of the resulting solutions are 5,10,15,20 & 25g/ml and the absorbance
were measured at 393nm and the calculation curve plotted.

Assay:

20 tablets were weighed accurately and the average weight was calculated and powdered.
150mg equivalent weight of the powder was weighed and dissolved in 100ml of 0.1N
NaOH.
From the above solution pipette out 1ml and dilute it to 10ml with 0.1N NaOH and
distilled water in1:1 ratio.

Category: Non-Steroidal anti-inflammatory.

Report: The percentage purity of Nimisulide by calibration curve method was found to
be____________.

Estimation of Diclofenac by UV-Visible

Spectroscopy
Aim: To estimate the percentage purity of diclofenac by UV-Visible Spectroscopy.
Apparatus: Volumetric flask, Curette, Pipette.
Chemicals: Diclofenac pure drug, Diclofenac tablets, 0.1N NaOH.
Principle: Diclofenac is a widely used analgesic and anti-pyretic and is estimated by UVSpectroscopy.
The principle involved in the absorption of radiation due to the presence of electrons in the
diclofenac that can be raised to higher energy levels by absorption of energy. The absorbance
was measured at 276nm and the calibration curve is constructed to determine the percentage
purity of the compound.

Procedure:
Preparation of standard dilutions:

100mg of diclofenac was weighed accurately and dissolved in little quantity of 0.1N
Noah and then diluted to 100ml with 0.1N NaOH.
10ml of the above solution was taken and diluted to 100ml with 0.1N NaOH.
From the above stock solution 5 standard dilutions are prepared by diluting 0.5ml, 1ml,
1.5ml, 2ml & 2.5ml with 0.1N NaOH to 10ml.
The concentrations that are obtained are 5,10,15,20 & 25g/ml and the absorbance were
measured by using 0.1N NaOH as blank.
The calibration curve was plotted between concentration and absorbance.

Assay:
Weigh and powder 20 tablets. Weigh accurately a quantity of powder containing about 50mg of
diclofenac sodium. Shake with 60ml of methanol in a 200ml volumetric flask and dilute it to
mark with methanol. Dilute 5ml of this solution to 100ml with methanol and measure the
absorbance of the resulting solution at the maximum at about 265nm.Calculate the content of
diclofenac from the absorbance.
Report: The percentage purity of diclofenac by calibration curve method was found to be
_________________

Estimation of Calcium Concentration by Flame

Photometry
Aim: To estimate the calcium concentration in the given sample
Apparatus: Volumetric flask, pipette
Principle: The technique of flame photometry is based on the measurement of emitted radiations
in a solution. When a solution of metal compound is introduced into the flame in the form of
droplets.
The fine residue forms the neutral atoms which absorb the energy from the flame & exited
and while relaxing there exited atoms emits radiations.The wavelengths of the emmited
radiations.The wavelengths of the emmited radiations is the characteristic of an element and can
be used foe quantitative analysis.while the intensity of emmited radiations denotes the
concentration of metal present in the sample.the alkali alkaline earth metals such as Na+,K+,Cr
+2,Ca+2, can be determined by flame photometry as the energy absorbed by the flame is quite
sufficient for the excitation of these elements . the quantitative results can be obtained by
plotting a calibration curve between the concentration and flame intensity.
Procedure: Prepare a series of standard solutions of 100ppm,200,300,400,500,600ppms
concentrations by taking sufficient amount of kcl and dilute upto 100ml by using distilled water.
Switch on the flame photometer and select the k+ filter. Set the gas in flame ,in order to get nonlumious flame &air pressure at 0.4-0.5cm2.Automize the flame intensity to 0.1 using distilled
water.Aotomize the flame intensity to 100% with weight concentration i.e, standard solution of
higher concentration .Measure the % flame intensity of all standard solutions &unknown sample.
Report: The concentration of calcium ions present in the given solution was found to be
______________ by flame photometry.

Estimation Of Sodium Concentrtion By

Flame Photometry

Aim: To determined the Sodium concentration in the given sample.

Apparatus: Measuring cylinder ,pipette ,volumetric flask.
Chemicals: Nacl, distilled water.
Principle: The technique of flame photometry is based on the measurement of emitted
radiations when a solution of metal compound is introduced into the flame, in the form of
droplets .the free residue forms the neutral atom which absorbs the energy from flame & exites
while relaxing these exited atoms emits the radiation. The Wavelength of the emitted radiation is
the characteristic of an element and can be used for quantitative analysis. While the intensity of
the emitted radiations denote the concentrations of the metal present in the solution. The alkali
and alkaline earth metals such as Na+
,K+,Cr+3,Cu+2, can be determined by flame photometry as the energy obtained by the flame is
quite sufficient for the excitation of elements . Flame photometry is also known as flame
emission spectroscopy because of the use of flame to provide an energy of excitation if atoms
introduced into the flame .
Flame photometry is a simple rapid method for the routine analysis of metals of I column of the
periodic table but it is also used for the certain transition elements such as copper, manganese,
the quantitative results can be obtained by plotting a calibration curve between the concentration
of the element & flame intensity.the process involved in the flame photometry was complex but
should follow the following steps.

Liquid sample containing the element is aspirated into the flme for formation of liquid
droplets .
Evaporation of liquid droplets results in formation of salt residue.
The decomposition of residue to the formation of free neutral atomswhich are unstable
quickly emits photons & return to the lower energy state.
The measurement of emission radiation in terms of wave length or intensity on basis of
flame photometry.

If E1&E2 represents the energy of higher &lower energy state ,the radiation emitted during the
changing energy levels may be defined by equation .

E2 - E1= h
h=planks constant

=Frequency of emitted light

=C /

hc
hc
= =
E 2E 1
We get E2-E1=
E2
Procedure: Prepare a series of standard solutions of 100ppm,200,300,400,500,600ppms
concentrations by taking sufficient amount of Nacl and dilute upto 100ml by using distilled
water. Switch on the flame photometer and select the sodium peak. Set the gas in flame ,in order
to get non-lumious flame & air pressure at 0.4-0.5 kg/cm2.Atomize the flame intensity to 100%
using standard solution of higher concentration. Measure the % flame intensity of all standard
solutions & unknown sample.
Report: The concentration of sodium ions present in the given solution was found to be
______________ by flame photometry.

Estimation Of Potassium Concentration by Flame

Photometry
Aim: To determine the potassium concentration in the given sample.
Apparatus: Volumetric flask, Pipettes.
Chemicals: KCl
Principle: The technique of flame photometry is based on the measurement of emitted radiations
in a solution. When solution of metal compound is introduced in to the flame in the form of
droplets, the solvent is evaporated and forms a fine residue of neutral atoms which absorb the
energy from the flame and exite. While relaxing these exited atoms emits radiations.
The wavelength of the emitted radiations is the characteristic of an element and can be used for
qualitative analysis. While the intensity of emitted radiations denotes the concentrations of metal,
present in the sample.
The alkali & alkaline earth metals such as Na+, K+, Ca2+ can be determined by flame photometry
as the energy absorbed by the flame is quite sufficient for the excitation of these elements
The quantitative results can be obtained by plotting a calibration curve between the concentration
and flame intensity.
Procedure:
Prepare a series of standard solutions of 100ppm, 200ppm, 300ppm, 400ppm, 500ppm, &
600ppm concentrations by taking specified amount of KCl and dilute it up to 100ml by using the
distilled water.
-

Switch on the flame photometer & select the potassium peak.

Set the gas in flame in order to get a non-luminous flame & air pressure at 0.4 to 0.5
Kg/m2. Atomize the flame intensity to 0% using the distilled water.
Atomize the flame to 100% using standard solution of higher concentration.
Measure the % flame intensity of all the concentrations & unknown sample.

Report: The absorbance of Potassium ion (K+) present in the given sample of KCl solution was
found to be __________ by flame photometry.

10

Identification Of Amino Acid By Ascending Paper

Chromatography
Aim: To develop a paper chromatogram or to perform paper chromatography by ascending
method.
Requirements: Wattman Filter paper, Spray gun.
Samples: Phenyl alanine & Cystine.
Chemicals: Ninhydrin, amino acid, Solvent (n-butanol:acetoc acid:water 4:1:5).
Preparation Of Solutions:
1. Solution A:
0.1% of
2. Solution B:
0.1% of
3. Ninhydrin solution: 0.2% (200mg was dissolved in 100ml of butanol).
Principle: Paper chromatography technique is a type of partition chromatography in which
substances are distributed between the two liquids i.e., stationary phase (usually water) which is
held in the fibers of the water in paper, the other is moving liquid or developing solvent and
called as mobile phase. The components of the mixture to be separated migrate at different rates
& appear as spot as different points on the paper.
This technique was used to separate mixture of organic substances such as dyes & amino
acids only. But now, this method has been perfected to separate cations and anions in organic
substances. In this technique, a drop of test solution is applied as spot on a wattmann filter paper
and the spot is dried. The paper is kept in a closed chamber and the edge of the filter paper is
dipped in to a solvent called developing solvent. As soon as the filter paper acts the liquid
through its capillary action and it reaches the spot of the test solution. The various substances are
moved by the solvent system at various speeds, when the solvent has moved these cations to a
suitable height the paper is dried and various spots are visualized by suitable reagent. The
movement of substances relative to the solvent is expressed in forms of Fro values.
Fro = distance travelled by solute from origin line
Distance travelled by solvent from origin
Procedure:
Take Wattmann filter paper, draw a thin line about 2 inches height from the bottom edge of the
paper. Draw mark on the paper should be in the direction of mobile phase. Mark two points
equidistant on a straight line and lebel them as A, B. Place the samples on the spots with a
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capillary tube. Hang the paper in a chromatographic chamber having the solvent. So that it
touches the surface of the solvent. Allow the solvent to run upto the 3/4 th of the paper. Remove
the chromatogram from the chamber and mark the solvent position with pencil. Dry the
chromatogram in air for 10 min and spray the dried chromatogram with Ninhydrin reagent. Dry
the chromatogram in an oven at 60C for 10-15 min. Purple colour spot will developed
components and the solvent front and compete the Fro values.
Report:
The Fro value of the given samples was found to be ______________ when the solvent system is
n-butanol: acetic acid: water 4:1:5.

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Identification of Amino Acid by Radial

Chromatography
Aim: To separate and identify amino acids by radial chromatography.
Preparation: Solution A - 0.1% L cosine
Solution B - 0.1% of Almandine
Solution C - Mixture of A & B solutions
Solvent Solution: n- butane: acetic acid: water (4:1:5)
Ninhydrine Solution: 0.2% (200mg in 100ml of butane)
Apparatus: Circular watts man filter paper, ninhydrine reagent, capillary tube
Principle: Radial chromatography is a type of paper chromatography which works on basic
principle. The solute is distributed between two immiscible liquids and helps in their separation
as it includes all liquids in the process. It is also a type of liquid - liquid chromatography. The
stationary phase is water i.e., based in the cellulosic fibers of the paper used in chromatography
and mobile phase is the solvent system solvated. After the development of chromatogram in the
radial technique, certain visualizing agents are used for the identification and for visualizing the
spots.
Procedure: In this technique a circular filter paper was employed. Then the sample which has to
be analyzed and placed on small circle which is drawn at the center of the radial paper through a
capillary tube.
Three points are marked on the circle which is drawn and label them as A, B, C. A drop of the
respective samples was placed on the spots that are marked. The paper is than fixed horizontally
on the Petridis in to the solvent and covered the paper by means of a lid. The solvent rises
through the wick and the solvent was allowed to run from front was marked with the pencil. Dry
the chromatogram in air for 10 min & sprayed with ninhydrine reagent. It was then dried at oven
at 60 c for 10- 15 min purple color spots were developed from the distance travelled by solute &
solvent front was measured.
Report: The Fro value of L- cosine & Leonine were found to be______________.

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Identification Of Amino Acid By Thin Layer

Chromatography
Aim: To separate and identify the amino acids by thin layer chromatography .
Apparatus: chromatographic technique , Glass plate ,Chromatograpic chamber ,Capillary tubes.
Chemicals: Ninhydrine ,n-butanol, Glacial acetic acid , Amino acids.
Preparation of solution: Solution A
Solution B -0.1
Solution c - 0.2%
Mobile Phase: n- butanol , acetic acid , water (4:1:5)
Principle: The principle involved in the TLC is adsorptionin this stationary phase is finely
divided adsorbent solid spread on a glass plte .The mobile phase is the liquid .
TLC is similar to paper chromatography except that thin (0.250nm) layer of some inert material
such as Al2o3 ,Mgo , Sio2 is used as asubstrate insteed of paper . A paper of one of these oxides
is made from a slurry of powder in a suitable inert solvent .
The slurry is poured evenly over a flate surface of the glass plate & dried. This plate couted with
slurry is placed in a suitable inert solvent .
The slurry is poured evenly over a flate surface of the glass plate and dried . This glass plate is
coated with slurry is placed in a suitable solvent system by spotting it with corresponding
sample.
The mobile phase solvent flows through the plate because of the capillary action against the
gravitational force .The components move acc to their affinities towards the adsorbent . The
component with more affinity towards stationary phase travells faster . This component are
separated on TLC plate based on affinity of components towards stationary phase.
TLC is mainly used for the depth and analysis of high molecular weight compounds and also
used for identification and separation of wide variety of mixtures , such as amino acids , dyes,
sugars, natural products etc.

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Procedure:
The prepared solvent system ( n-butanol : acetic acid : water in 4:1:5) was kept
aside for saturation of the chamber .
The redymade pre- coated plates were taken on which a straight line was drawn
above 2 inches from the edge of TLC plate.
Two points were marked equidistant on the straight line which were labelled as
A& B.
The TLC plate was placed in the saturated chamber for the development of the
chromatogram.
The spots were visvalated by spraying ninhydrin reagent followed by drying and
the florescent sample was observed under up- spectroscopy.
Report: The Fro value of the given sample was found to be______________.

15

Estimation of paracetamol by HPLC

Aim: To estimate the amount of paracetamol by HPLC
Apparatus: Volumetric flask, beaker, pipette
Chemicals: Paracetamol standard, sample, methanol for HPLC, water for HPLC
Principle: The principle is adsorption. In isocratic HPanalyzelyte is forced through a column of
stationary phase by pumping a liquid (mobile) phase at high pressure through a column. The
sample to be analyzed is introduced in a small volume stream of mobile phase and is retained by
specific chemical/ physical interface with stationary phase as it travels the length of column,
amount of retardation depends on the nature of the analyze, stationary and mobile phase
composition. The time at which a specific analyze is eluted is called Retention Time and is
considered as a characteristic of given analyze.
The use of pressure increases the linear velocity giving the component less time to differ within
column leading to improved resolution in the resulting chromatogram.
Common solvents used include miscible combinations of water/various organic liquids. Water
may contain buffer/salts to assist in separation of analyze components such as it acts as an ion
pairing agent.
Procedure:
Preparation of mobile phase: HPLC grade methanol was taken, filtered under vacuum, and
finally it was solicited for 15 min to remove dissolved gases

100 mg of drug was dissolved in 100 ml of methanol to give a 1 mg/ml solution

From the above solution, 0.1 ml, 0.2 ml, 0.3 ml, 0.4 ml and 0.5 ml were taken and
volumes diluted to 10 ml to give 10, 20, 30, 40, 50 g/ml respectively
The solutions were filtered and solicited to remove dissolved gases.
The resulting solutions were injected into HPLC columns and RFs were found. The
calibration curve was constructed using area under curve vs. concentration.

Report: The concentration of given sample of paracetamol was found to be ______________.

16

Estimation of caffeine by HPLC

Aim: To estimate the amount of caffeine in the given sample by HPLC
Apparatus: Volumetric flask, beaker, pipette
Chemicals: caffeine pure, sample, methanol for HPLC, HPLC grade distilled water
Principle: The principle is adsorption. In isocratic HPLC, analyze is forced through a column of
stationary phase (a tube packed with small bound particles with certain surface chemistry) by
pumping a liquid (mobile) phase at high pressure through a column. The sample to be analyzed is
introduced in a small volume stream of mobile phase and is retained by specific chemical/
physical interface with stationary phase as it travels the length of column, amount of retardation
depends on the nature of the analyze, stationary and mobile phase composition. The time at
which a specific analyze is eluted is called Retention Time and is considered as a characteristic
of given analyze.
The use of pressure increases the linear velocity giving the component less time to differ within
column leading to improved resolution in the resulting chromatogram.
Common solvents used include miscible combinations of water/various organic liquids. Water
may contain buffer/salts to assist in separation of analyze components such as it acts as an ion
pairing agent.
Procedure:
Preparation of mobile phase: HPLC grade methanol was taken, filtered under vacuum, and
finally it was solicited for 15 min to remove dissolved gases

100 mg of drug was dissolved in 100 ml of methanol to give a 1 mg/ml solution

From the above solution, 0.1 ml, 0.2 ml, 0.3 ml, 0.4 ml and 0.5 ml were taken and
volumes diluted to 10 ml to give 10, 20, 30, 40, 50 g/ml respectively
The solutions were filtered and solicited to remove dissolved gases.
The resulting solutions were injected into HPLC columns and RFs were found. The
calibration curve was constructed using area under curve vs. concentration.

Report: The concentration of given sample of caffeine was found to be_____________.

17

Interpretation of IR-Spectrum
Aim: To interpret the IR-Spectrum of 3-(N, N-diethyl amino) ethyl-2-thio-6-bromoquinazolin-4(3H) one.
Instrument: IR Spectrometer, Alpha-T-Broken.
Principle: IR Spectrum is the result absorption of light by vibrating molecules. In any
molecule it is known that atoms or groups of atoms are connected by bonds. These bonds
are analogues to springs and not rigid in nature. Because of continuous motion of the
molecule the maintain some vibrations with some frequency, characteristic to every
portion of the molecule. This is called natural frequency of vibration. When the energy in
the form of IR is applied and when applied IR frequency equal to natural frequency of
vibration absorption of IR taken place and peak is observed.
Procedure: Functional group analysis of the IR spectrum in Ker pellets. Here in the
spectrum we found NH functional group at 3378.89cm-1 as stretching vibrations.
We found in this spectrum, CH- aromatic group at 3185.24cm-1 as stretching vibration.
There is a C=O group found at 1671.43cm-1 as a stretching vibration.
The given spectra consist of a C-H group at 824.35cm-1 as a bending vibration.
Report:

Interpretation of Mass Spectroscopy of Drug

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Aim: To interpret the mass spectroscopy of given drug.

Instrument: Mass spectrometer.
Principle: Spectroscopy is the branch of science dealing with the study of interaction of
electromagnetic radiation with matter. In the simplest mass spectrometer organic
molecules are bombarded with electrons and converted to highly energetic positively
charged ions. The loss of an electron from a molecule leads to a radial action and we can
represent this process as mom+. The molecule ion m+ commonly decomposes to a pair of
fragment and may be radicals or anion or a small molecule plus an action. Thus,
m+ m1+ + m2* (or) m1- + m2*
Procedure: Collisions induced dissociation spectra was acquired in the positive ion on a
moss (skied) ARI-4000 triple quadruple mass spectrometer with direct infusion of each
acetone at a concentration of 10m in 50% methanol at a flow rate of 25l/min. The
instrument was operated with spray voltage with of 5.5Kv a dechistering potential of 5ev
a source temperature of 100 C a G 8i value of the certain gas set at 10
Results & Discussion:

Interpretation of 13C-NMR of Drug

19

Aim: To interpret the 13C-NMR of given drug

Apparatus: NMR
Principle: Spectroscopy is a branch of science dealing with study of interaction of
electromagnetic radiation with matter. The most important consequences of such
interaction are discrete amount call quanta. NMR is concerned with the magnetic
resonance of certain atomic nuclei. Notably the nucleus of the hydrogen atom, the proton
so that of the C-13 isotope of carbon studying a molecule by NMR spectroscopy enables
us to record difference in the magnetic properties as the various magnet nuclei present
Procedure:

The molecular formulae of the given drug which contains H atoms in 4- chemical

environments, so that we can inapt the 4 peaks.

The aromatic peak resonates at 7.17 because of anisotropic effect as multiplexes.
The N atom present at 10th position & 5 member ring, so the H atoms

experiencing the electro negativity & anisotropic effect, so they resonate at 3.92.
The H of COOH group present at 2 nd position resonates at 10.48 due to the EN of

oxygen located at the adjacent carbon.

The remaining H at HC and HC are in same chemical environment they resonate
at 2.49 & experience doublet

Report:

Interpretation of H-NMR of Drug

Aim: To interpret the H-NMR of given drug
Apparatus: NMR, DMSO-do

20

Principle: Spectroscopy is the branch of science dealing with study of interaction of

electromagnetic radiation with matter. The most important consequences of such
interaction are that energy absorbed as emitted by the matter is discrete amount called
quanta. NMR is concerned with the magnetic resonance of certain atomic nuclei, notably
the nucleus of the H atom & proton. So that of the C-13 isotope of carbon studying a
molecule by NMR spectroscopy enables us to record difference in the magnetic
properties as the various magnetic nuclei present.
Procedure:
the compound contains H atoms in 4- chemical environments, so that we can expect 4
peaks

the aromatic peak resonate at 7.17 because of anisotropic effect as multiples.

The N-atom present at 10th position and 5 member ring, so the H atom experiencing the
electro negativity atom and anisotropic effect so they resonate at 3.22.
The H of COOH group present at 2nd position resonates at 10.48 due to the EN of
oxygen located at the adjacent carbon singlet is shifted to down field.
Report:

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