abhi final

Analysis of active pharmaceutical ingredient levetiracetum drug by HPLC and GC method.
INTRODUCTION
A drug is a chemical used medicinally for treating diseases and injuries. Many drugs or
medicines are entirely legal, readily available and are sold as over the counter medications
without the need for a prescription. Over the counter drug include categories of drugs such as
pain relievers, Antacid, Alcohols, Caffeine, and Vitamins. Dosage information and
precaution are strictly followed in order to prevent accidental injury or harm.
Levetiracetam is an anticonvulsant medication used to treat epilepsy

Chromatography is a technique widely used to separate the mixture of compounds into its
individual form and obtain pure compounds from the mixture.
Principle:- chromatography is based on the principle of separation of compounds into
different bands and the identification of those bands. The preferential separation is done due
to different affinities of compounds towards stationary and mobile phase.
Components:-
 Stationary phase:- The stationary phase is one which stays motionless and allow
sample to move over it.
 Mobile phase:- This is the chromatographic liquid phase and it helps the sample
move over the stationary phase.
Types of chromatography:-
1. Thin layer chromatography

2. Paper chromatography
3. Column chromatography
4. Ion exchange chromatography
5. Gas chromatography
6. High performance liquid chromatography
High performance liquid chromatography is a technique in analytical chemistry used to

separate, identify and quantify each component in a mixture.
Gas chromatography refers to a physical process by which a mixture is separated into its
constituents by moving gas phase passing over a stationary sorbent.
Karl-Fischer Titration is classic titration method in analytical chemistry that was colorimetric
or volumetric titration to determine amount of water in a sample.
Department of Chemistry. Government Science College Chitradurga. Page 1

LITERATURE SURVEY
Literature survey on chromatographic technique reveals that they are highly useful and
effective analytical methods for the analysis of drugs quantitatively as well as qualitatively.
These methods are especially revealed by the famous
 Journal of chromatography A. 1036, Gerber, F; Krummen, M; Potgeter, H; Roth, A;
Siffrin, C ; Spoendlin, C2: 127-133 (2004).
 Journal of chemical education , 74 : 45 by Karger, Barry L. (1997).
These technique of separation and identification of newly synthesized drugs
promoted us to take up this project

OBJECTIVES
The main objective of this project is to study the parameters of drugs composition and
to get satisfactory result and to make the method to be rapid, simple, accurate, linear,
selective, economical, and reproducible and to have short run time and to achieve low cost
technology which makes this method economically alternative for most clinical laboratories.
PLAN OF THE WORK
Analysis of chemicals by chromatographic method is one of the important and
accurate methods of all the techniques. So this has attracted much interest due to their wide
range of usefulness. In the last few years the method is developed as a new challenge to
chemical and pharmaceutical industries for the analysis of different chemicals and drugs. In
addition the study and development of technology by chromatography could be helpful in the
design and development of new methods in finding the new parameters.
The main aim of this project is to study the parameters of drug components to attain its
effective function.

INTRUDCTION TO LEVETIRACETUM
 Levetiracetum is a new and novel antiepileptic drug ( AED) with an interesting
history and unique properties, and it may be reprehensive of a new class of AEDs.
 The history of effective drug began with the intrudction of the bromides in 1857 based
on later discredited theories of the cause of epilepsy 1.Phenobarbital(PB) was
introduced in 1912 for the treatment of epilepsy because of its sedative properties1.
 Levetiracetum is an anticonvolusant medication used to treat epilepsy.
 Levetiracetum may selectively prevent hyper synchronization of epileptiform burst
firing and propagation of seizure activity.Levetiracetum binds to the synaptic vesicle
protein SV2A,which is thought to be involved in the regulation of vesicle exocytose.
Although the molecular significance of levetiracetum binding to synaptic vesicle
protein SV2A is not understood, levetiracetum and related analogs showed a rank
order of affinity for SV2A which correlated with the potency of their antiseizure
activity in autogenic seizure prone mice.
STRUCTURE

CHARACTERISTICS OF LEVETIRACETUM
GENERIC NAME : LEVETIRACETUM
IUPAC NAME : (2S)-2-(2Oxopyrrolidin-1-yi)butadiene.
TRADE NAME : Diovan
APPEARANCE : Coloured crystals
SOLUBILITY : Ethanol, methanol, water, freely soluble in chloroform and
sparingly soluble in Acetonitrile.
MOLECULAR FORMULAE : C8H14 N2 O2
MELTING POINT : 1070 C
MOLAR MASS : 170.212g/mol.
MEDICAL USE
 LEVETIRACETUM has been approved in the United states as add on treatment for
partial , myoclonic,and tonic clonic seizures.
 Levetiracetum has potential benefits for other psychiatric and neurologic conditions
such as Tourette syndrome ,anxiety disorder, and Alzhemeirs disease. However ,its
most serious adverse effects are behavioral,and its benefit risk ratio in these
conditions is not well understood.

 Levetiracetum has not been found to be useful for treatment of neuropathic pain , nor
for treatment of essential tremors.Levetiracetum has not been found to be useful for
treating autism ,but is an effective treatment for partial,myoclonic,or tonic –clonic
seizures associated with autism spectrum disorder.
 Levetiracetum is a pregnancy category C drug.Studies in female pregnant rats have
shown minor fetal skeletal abnormalities when given maximum recommended human
doses of levetiracetum orally throughout pregnancy and lactation.
MECHANISM ACTION OF DRUG

The precise mechanism by which levetiracetum exerts its antiepileptic effect is unknown.
However the drug binds to SV2A, a synaptic vesicle glycoprotein, and inhibits presynaptic
calcium channels,reducing neurotransmitter released acting as a neuromodulator.This is
believed to impede impulse conduction across synapses.
SIDE EFECTS
All drugs may cause side effects ,However many people have no side effects or only have minor side
effects.
 Loose stools
 Dizziness
 Feeling sleepy
 Stuffy nose
 Nose and throat irritation
 Belly pain
 Not able to sleep
 Feeling tired or weak
 Headche
 Not hungry
 Upset stomach or throwing up.

DATA ANALYSIS OF LEVITERACETUM BY HIGH PERFORMANCE

LIQUID CHROMATOGRAPHY METHOD:-
Introduction
High performance liquid chromatography is a technique in analytical chemistry used to

separate , identify each component in a mixture. It is an effective type of column
chromatography which is widely used in pharmaceuticals it is very useful to determine the
assay and related substance in drug. In general HPLC is used to separate the components of a
mixed drug substances.
In HPLC column plays a significant role in separation of different compounds because it

contains stationary phase. Stationary phase is a bed of polar and non-polar particles according
to type of column. Polar and non-polar compounds are used according to the nature of the
sample to be analyzed.
PRINCIPLE
In HPLC, eluent from the solvent reservoir is filtered pressurized and pumped through the
column. A mixture of solutes injected at the top of the column is separated into components.
Individual solutes are monitored by the detector and recorded automatically.
Types of HPLC based on mode of separation:-
 Normal phase chromatography:-

The stationary bed is strongly polar in nature (eg:silica gel) and the mobile phase is non
polar (such as n-hexane or Tetrahydrofuran). Polar substances are thus retained on the polar
surface of the column packing longer than less polar materials.
 Reversed phase chromatography:-

The stationary bed is non polar(hydrophobic) in nature, while the mobile phase is a polar
liquid, such as mixtures of water and methanol or Acetonitrile. Here the more non polar the
material is longer it will be retained.

Characteristics feature of HPLC
 High resolving power and speedy separation.

 Accurate quantitative measurements.
 Continuous monitoring of column effluent.
 It can determine multiple component in a single analysis.
 Separated components can be easily collected from the mobile phase for further
characterization.
 Rapid technique applicable to all kinds of chromatgraphic methods such as
absorption, ion exchange, partition and exclusion chromatography.
 A variety of solvents sample recovery is easy.
 Analyses are completed it in few minutes with excellent Precision and accuracy.
 HPLC provided with automatic analytical procedure and data handling
 Repetitive and reproducible analysis using the same column
 Chromatography separation in HPLC is the result of specific interaction between the
sample molecules with both the stationary phase and mobile phase.
INSTRUMENTATION

Schematic representation of HPLC:-
A typical modern liquid chromatography consist of following components
 Solvent delivary system

 Sample injection system
 Column
 Detector
 Recorded
 Data control and display
Solvent delivary system
The pump is one of the most important components of HPLC, since its performance
directly affects retention time, reproducibility and detector sensitivity. The pump delivers a
steady steam of solvent from the reservoir to detector through the column. The Pump can
deliver solvent at a pressure up to 10000 psi with flow rate over 50 cm3 per minute. Most of
the separations done by HPLC require pressure between 400 and 1500 psi.
Types of pumps
 Gas displacement pumps

 Pneumatic pump
 Syringe pump
 Reciprocating pump

 Sample injection system

Sample injection system consisting of two types of injecting system
 Syringe injection
 Stop-flow injection
HPLC VIALS
 Column
Columns are considered as the ”HEART OF CHROMATOGRAPHY”. The column is

made from precision bore polished stainless steel tubing , typical dimensions being 10
cm-30cm long and 4mm-5mm in internal diameter. Temperature of the column affects
speed of affinity, viscosity of the solvent, diffusion and solubility of the sample. Columns
are thus used in thermostatic ovens.
 Detector:-

In HPLC , the function of detector is to monitor the mobile phase as it emerges from
the column.
Characteristic of a detector
 Sensitivity
 Linear response
 Type of response
Types of detector
 Ultraviolet detector
 Photodiode array detector
 Ultraviolet detector
The UV absorption detectors have been used widely in HPLC. It is on the principle
of absorption of UV visible light as the effluent from the column is passed through a
flow cell held in radiation beam.
 Photodiode array detector:-

Can be recorded whole of the spectrum many times a second. In these detector,
polychromatic light is passed through the HPLC flow cell and the emerging radiation
is diffracted by a grating to fall on the array.
 Recorder
The signal from the detector is recorded as deviation from a base line. Two open
recorders are used with instruments having two detectors. The peak position along the
curve relative to the starting point, denotes the particular components with proper
calibration , the height or area of peak is a measure of the amount of the component
present in the sample.

Uses of HPLC:-
This technique is used,
 In chemistry and biochemistry research analysing the complex mixtures.
 Purifying the chemical compounds.
 Developing processes from synthesizing chemical compounds.
 Isolating natural products and predicting the physical properties.
 It is also used in a quality control to ensure the purity of raw materials.
 To control and improve process yield.
 To quantity assays of final products.
 To evaluate products stability and monitor degradation.

DATA ANALYSIS OF LEVETIRACETUM BY HIGH PERFORMANCE

LIQUID CHROMATOGRAPHY
Before data analysis of LEVETIRACETUM by HPLC first we have to prepare following
solutions.
Preparation of Buffer solution:
Weigh accurately about 0.26g of water and mix well using a magnetic stir bar until the
material is completely dissolved. Check the pH an adjust to 5.50±0.05 width 2% aqueous
solution of potassium hydroxide. Filter the solution through a 0.45 µm nylon membrane and
degas by sonicating for 5 minutes.
Preparation of Mobile Phase A:
Transfer 950ml of Buffer solution and 50ml of acetonitrile into 1L bottle mix well using a
magnetic stir bar. Filter the solution through a 0.45µm nylon membrane and degas by
sonicating for 5 minutes.
Preparation of Mobile Phase B:
Use filtered and degassed acetonitrile as Mobile Phase B.
Preparation of Diluent:
Use Mobile phase A as a diluent.
Preparation of System suitability solution:

Weigh accurately about 5mg of LVT standard and transfer into 25ml volumetric flask.
Dissolve in 2.5ml of 0.1N Pottassium hydroxide.Keep the mixture at room temperature for a
about 15 min and then neutralize by adding 25ml of 0.1N hydrochloric acid.Weigh about
2mg of LTM standard to it.Dissolve and dilute to volume with diluents.
Preparation of Sample solution:
Weigh accurately about 125mg of sample in duplicate and transfer into two separate 25ml
volumetric flask. Dissolve and dilute to volume with diluents. Lable these solution as sample
solution -1, and sample solution -2.
Preparation of diluted standard solution:
Weigh accurately about 5mg of LTM standard in 10 ml volumetric flask dissolve and diluted
to volume with diluent. Transfer 1ml of these solution into 100ml volumetric flask dilute to
volume with diluents.
HPLC Chromatographic condition:

Column YMC Pack OPC .AQ(150×4.6) 3µm equivalent
detector UV-Visible detector
Wavelength of detector 205nm
Flow rate 0.9ml/min
Column temperature 250C
Injection volume 10µm
Runtime 40min
Relative retention time 2-pyridinal RRT about 0.43
LTMacid RRT 0.5g
Levetiracetum RRT 1.00( RT about 16.2m)
LTM 1,RRT about 1.2 sec.

HPLC Analysis:
 Equilibrate the HPLC system with mobile phase though the column until a steady
baseline is obtained.
 Condition the column by running gradient program given
 After the system has equilibrated,make one junction of diluents blank.
 Make one injection of system suitability solution
 Make three injections of diluted standard solution
 Make one injections of system suitability solution at the end of analysis.
SYSTEM SUITABILITY CRITIRIA:
 The relative standard deviation for standard solution is more than 2%.
 Resolution between Levetiracetum and LTM 1 peaks should be more than 7.0 in
chromatogram obtained with system suitability solution.

GRAPHS



DATA ANALYSIS OF LEVETIRACETUM BY GC

INTRODUCTION
 Modern gas chromatography (GC) was invented by MARTIN and JAMES IN
1952 and has become one of the most important and widely applied analytical
techniques in modern chemistry.
 GRIFFIN and GEORGE probably manufactured the first commercial GC system
in 1954.
Instrumentation of GC
Schematic representation of GC

High pressure cylinder containing a carrier gas
 Sample injection system

 Column
 Thermal compartment and the
 Detector
 Supply of carrier gas from high pressure cylinder

The carrier gas is either H2 , He ,CO2 or Ar the choice depends on factors such as
nature of sample , purity, availability, consumption , column efficiency and the type of
detector employed. Thus helium is preferred when thermal conductivity detector are
employed because of its high thermal conductivity relative to the vapours of most
organic compounds. Flow rate greatly influence column efficiency.
 Injection system used in capillary GC

 Split injection method
 Split less injection system
 Direct on column method
 Automatic sampler
 Headspace sampling

Split injector:-
 Column
The actual separation of sample component are effected in the column. The
nature of the solid support, the type and amount of liquid phase, the method of
packing ,length and temperature are important factors in obtaining the desired
resolution. The Column is enclosed in thermostatically controlled oven so that its
temperature is held constant, ensuring reproducible conditions. The operating
temperature may range from ambient over 673 K.
Types of column
 Packed column
 Capillary column
There are two types of columns: OPEN TUBULAR COLUMN and PACKED COLUMN
 Open tubular columns are also known as capillary columns

 Packed columns are made of glass or metal tubing which is densely packed
with a solid support like diatomaceous earth.
 Thermal compartments

The column is operated thermally if a sample contains several compounds having a

wide range of boiling points. Now linear and non- linear temperature of the column
can be maintained uniform about the 273.1 k by the use of vapour jacket or liquid
baths.
 Detector
A detector located at the exit of the separation column senses the presence of small
amounts of individual components as they leave the column.
The choice of detector depends on factor such as the concentration level to be

measured and the nature of the separated components.
Types of detector
Ionization detector
Carrier gases behave as perfect insulator at normal temperature and pressure. The
increased conductivity due to charge ions in the effluent from the column provides high
sensitivity which is a feature of ionization based detector. Thus ID operates by measuring the
electrical conductivity of a gas which is directly proportional to the concentration of the
charged particles within the gas . Current ionization detector thermionic ionization detector ,
photo ionization detector and electron capture detector.
Flame ionization detector

The basis of FID is that the effluent from the column is mixed with H 2 and burned in air to
produce a flame which ionizes the solute molecules having ionization potential. The burner
jet is the negative electrode while the anode is usually a wire extending into the tip of the
flame. When ionizable material from the column effluent enters the flame and is burned, the
current markedly increases. The current flows through an external resistor is sensed as a
voltage drop, amplified and finally sent to an output device, a recorder.
Schematic diagram of a flame ionization detector
Applications of Gas chromatography

 Gas chromatography is used in many different fields such as pharmaceuticals
cosmetics and Evan environmental toxins.
 Air samples can be analysed using gas chromatography. Most of the time air quality
control units used gas chromatography coupled with flame ionization detector in order
to determine the components of a given air sample.
 Gas chromatography is another useful method which can determine the components
of a given mixture using the retention time and the abundance of the samples.
 The precission the accuracy of the method is very high.

 The technique has strong separation power and even Complex mixtures can be
resolved into constituents.
 In industry:
 Ultrasensitive detector in GC are useful in the field of food products.
 Fatty acids, steroids, high boiling and thermally unstable materials have been
separated after converting them into the volatile and thermally stable
derivatives such as esters, acetates or trifluroacetates.
 A large number of industrial products including herbicides, pesticides,
fertilizers, pharmaceuticals, cosmetics, perfumes, protective coatings, plastic
materials, alcoholic beverages, rubber and rubber products , soap and synthetic
detergents have been analysed and separated by gas chromatography. The
technique is used in determination of water in butane gas , creams, emulsion ,
ointments and pastes etc. crude petroleum products gasoline, hydrocarbons, N
and S compounds have been separated by GC and identified by IR , NMR ,
UV and MS techniques.
 In Elemental analysis:
 In determination of C, H and N.
 In determination of Sulphur.
 In determination of total organic carbon.
DATA ANALYSIS BY LEVETIRACETUM GAS CHROMATOGRAPHY

Before data analysis of levetiracetum by gas chromatography we have to prepare following
solutions.
Preparation of stock solution:
Transfer accurately about 500mg of ethyl acetate ,500mg of acetone ,500mg of ethanol and
60 mg of Dichloro methane into 100mL volumetric flask containing 50Ml of dimethyle
sulphoxide make upto the mark and transfer 1ml of this solution into 20ml volumetric flask.

Preparation of Sample solution:
Weigh accurately about 100mg of sample into two separate 20ml headspace vials.Add 2ml of
dimethyle sulphoxide. Close the vial with a septum and seal with an aluminium crimp cap.
Preparation of Standard solution:
Transfer 2ml of standard stock solution separately into required number of 20ml headspace
vials and close the vial with a septum and seal with aluminium crimp caps.
Preparation of Blank;
Transfer 2ml of dimethylsulphoxide into 20ml headspace vial. Close the vial with a septum
and seal with an aluminium crimp cap.
GC CONDITION:
1m
Column SPB -624m supeico,60m
length ,320mminternal diameter and 1.8µm
film thickness or equivalent
Carrier column nitrogen
Flow rate 105ml /min
Detector temperature 2500c
Injector temperature 1700C
Split ratio 1:10
Oven temperature Initial 400C hold for 22min ,increases at

300C /MIN TO 2200C .hold at 2200C FOR 8
min
Relative retension time Ethanol ,RRT about 1.00
Acetone ,RRT about 1.17
Dichloroethane RRT about 1.39
Ethyleacetate RRT about 2.20
Headspace Condition:
Oven equilibrium temperature 1000C

Loop temperature 1050C
Transfer line temperature 1100C
Oven equilibrium time 15 min
Pressurization time 0.5 min
GC Analysis:
 Allow the GC to equilibrate at 400C until a steady baseline is obtained.

 After the system has equilibrated , make one injection of blank preparation.
 Make one injection each from six standard vials, ensure that system suitability
parameter meets the specified criteria.
 Make one injection each from two sample preparation.
 Make one injection of std preparation at end of the sequence or after every 5 th batch.

System Suitablity criteria :
Percentage relative standard deviation of peak areas for six replicate injection of standard
solutions for each solvent should be less than 10
Resolutions between ethylacetate and cyclohexane peaks obtained from first injections of
standard solutions should be more than 5.
SPECTRUM OF GC BLANK
SPECTRUM OF GC SAMPLE

SPECTRUM OF GC STANDARD
CALCULATION AND RESULT:

Calculate the solvent in the indivisual sample preparation using the following formulae and
report the average result.
=
Area of solvent of interest ∈the test Solution Wt of the Std Solvent 5 1 6
X X X X 10
Average area of Solvent of interest ∈ Standrad Solution 100 25 Wt of the Sample
At Ws 5 1 6
Concentration in PPM = X X X X 10
As 100 25 Wt
Where, At is the area of solvents peak in test standard sample solution.
As is the mean area of respective solvent peak in six standard solutions.
Ws is the weight of respective solvents reference / working standard in mg.
Wt is the weight of the sample is mg.
95.98 500 5 1 6
= X X X X 10
38.50 100 25 100
= 24,298 PPM

DETERMINATION OF MOISTURE CONTENT IN SAMPLE
Karl-Fischer Titration
Karl-Fischer Titration is a classic titration method in analytical chemistry that was
colorimetric or volumetric titration to determine amount of water in sample. It was invented
in 1935 by German chemist Karl Fischer.
INSTRUMENT
Apparatus:-
 Karl-fisher titrator
 Analytical balance Sensitivity 0.1g
 Closed glass weighing spoon.

Scope
This method may be used for any kind of dried milk products especially those containing
crystallized lactose.
PRINCIPLE
The determination of total moisture by Karl Fischer titration is a calculation based on the
concentration of iodine in the karl-fisher titrating reagent consumed in the titration. The end
point of the titration is determined by dead stop end point method.
Reagents
 Karl Fischer reagent

 Methanol and purified water
PROCEDURE
Transfer about 25 ml of methanol into Karl Fischer titrate vessel and titrate with Karl
Fischer reagent to neutralize determine the end point potentiometrically. Weigh accurately
about 2 g of sample, transfer into the Karl Fischer titration vessel. Titrate with Karl Fischer
reagent. Calculate the strength of Karl Fischer reagent as water equivalence (mg/ ml) using
the formula.
Weig h t of sample taken∈grams ×100

Water equivalence =
Volume of KF reagent consumed
Determination of water in sample

Weigh accurately quantity of material specified in relevant method of analysis and
transfer into a titration vessel containing previously titrated methanol with Karl Fischer
reagent.
Titrate with Karl Fischer reagent and determine the end point potentiometrically in duplicate
and note down the volume of Karl Fischer reagent consumed.
Titer value ×Water equivalence × 100

Water content ( % )=
1000 ×Weig ht of sample

Finally calculate the amount of water in the sample by using the following
formula.
Titer Value X K . F . factor X 0 .1

Water Content (%) = Weight of Sample
= 0.19%

RESULT AND DISCUSSION :
The following table consists of test performed on the sample along their result and specifications.
SL TEST SPECIFICATIONS RESULTS

No
1 Description White crystalline powder White crystalline
Taste bitter powder
2 Identification RT of the principle peak in the Complies
By HPLC chromatogram of the sample
By GC corresponds to that in the
chromatogram of the reference
standard , as obtained in chiral purity
method
3 Solublity Very soluble in water(104g/100ml) Meets the requirements
freely soluble in
chloroform(65.3/100ml)
Methanol(53.6g ml)
Ethanol(16.5g/ml)
Sparingly soluble in acetonitrile and
partially soluble in n-hexane.
4 Melting point 1170 C 1170 C
5 Water content by KF Not more than 0.05% 0.19 %
6 Identification by HPLC The RT of the major peak for Complies

valsartanfron sample solution should
corresponds to that of valsartan
enantiomer from the system
suitability solution.

CONCLUSION:-
Different set of analysis method is being developed for the analysis of bio chemical
compounds such as drugs; LEVITERACETUM is one which plays an important role in
analysis process.Material and active pharmaceutical ingredients are tested before they are
used for the manufacturing in large scale and based on their results obtained products are
passed with the Analytical Development Lab (ADL). Statistical analysis of the results shows
that the proposed procedure has good precision and accuracy. And it shows satisfactory
results for all the method validation parameters tested. LEVITERACETUM is used to treat
epilepsy drug. According to the results obtained from the different tests it shows that values
are with of pharmacopeia specification procedure so these products can be produced in large
scale and may be employed for routine quality control analysis.

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Analysis of active pharmaceutical ingredient levetiracetum drug by HPLC and GC method.

Levetiracetam is an anticonvulsant medication used to treat epilepsy

1. Thin layer chromatography

High performance liquid chromatography is a technique in analytical chemistry used to

Department of Chemistry. Government Science College Chitradurga. Page 1

These methods are especially revealed by the famous

 Journal of chromatography A. 1036, Gerber, F; Krummen, M; Potgeter, H; Roth, A;

Siffrin, C ; Spoendlin, C2: 127-133 (2004).

 Journal of chemical education , 74 : 45 by Karger, Barry L. (1997).

These technique of separation and identification of newly synthesized drugs

promoted us to take up this project

Department of Chemistry. Government Science College Chitradurga. Page 2

PLAN OF THE WORK

Analysis of chemicals by chromatographic method is one of the important and

design and development of new methods in finding the new parameters.

Department of Chemistry. Government Science College Chitradurga. Page 3

 Levetiracetum is a new and novel antiepileptic drug ( AED) with an interesting

on later discredited theories of the cause of epilepsy 1.Phenobarbital(PB) was

 Levetiracetum is an anticonvolusant medication used to treat epilepsy.

 Levetiracetum may selectively prevent hyper synchronization of epileptiform burst

firing and propagation of seizure activity.Levetiracetum binds to the synaptic vesicle

protein SV2A,which is thought to be involved in the regulation of vesicle exocytose.

Although the molecular significance of levetiracetum binding to synaptic vesicle

activity in autogenic seizure prone mice.

Department of Chemistry. Government Science College Chitradurga. Page 4

IUPAC NAME : (2S)-2-(2Oxopyrrolidin-1-yi)butadiene.

TRADE NAME : Diovan

APPEARANCE : Coloured crystals

SOLUBILITY : Ethanol, methanol, water, freely soluble in chloroform and

sparingly soluble in Acetonitrile.

MOLECULAR FORMULAE : C8H14 N2 O2

MELTING POINT : 1070 C

MOLAR MASS : 170.212g/mol.

partial , myoclonic,and tonic clonic seizures.

conditions is not well understood.

Department of Chemistry. Government Science College Chitradurga. Page 5

treating autism ,but is an effective treatment for partial,myoclonic,or tonic –clonic

seizures associated with autism spectrum disorder.

 Levetiracetum is a pregnancy category C drug.Studies in female pregnant rats have

doses of levetiracetum orally throughout pregnancy and lactation.

MECHANISM ACTION OF DRUG

calcium channels,reducing neurotransmitter released acting as a neuromodulator.This is

believed to impede impulse conduction across synapses.

Department of Chemistry. Government Science College Chitradurga. Page 6

DATA ANALYSIS OF LEVITERACETUM BY HIGH PERFORMANCE

High performance liquid chromatography is a technique in analytical chemistry used to

In HPLC column plays a significant role in separation of different compounds because it

Types of HPLC based on mode of separation:-

 Normal phase chromatography:-

 Reversed phase chromatography:-

Department of Chemistry. Government Science College Chitradurga. Page 7

Characteristics feature of HPLC

 High resolving power and speedy separation.

Department of Chemistry. Government Science College Chitradurga. Page 8

Schematic representation of HPLC:-

A typical modern liquid chromatography consist of following components

 Solvent delivary system

Solvent delivary system

 Gas displacement pumps